WO2013159108A2 - Oligomeric compounds comprising bicyclic nucleotides and uses thereof - Google Patents

Oligomeric compounds comprising bicyclic nucleotides and uses thereof Download PDF

Info

Publication number
WO2013159108A2
WO2013159108A2 PCT/US2013/037638 US2013037638W WO2013159108A2 WO 2013159108 A2 WO2013159108 A2 WO 2013159108A2 US 2013037638 W US2013037638 W US 2013037638W WO 2013159108 A2 WO2013159108 A2 WO 2013159108A2
Authority
WO
WIPO (PCT)
Prior art keywords
comprises
moe
nucleoside
certain embodiments
deoxynucleosides
Prior art date
Application number
PCT/US2013/037638
Other languages
French (fr)
Other versions
WO2013159108A3 (en
Inventor
Susan M. Freier
Eric E. Swayze
Original Assignee
Isis Pharmaceuticals, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to US201261636513P priority Critical
Priority to US61/636,513 priority
Application filed by Isis Pharmaceuticals, Inc. filed Critical Isis Pharmaceuticals, Inc.
Publication of WO2013159108A2 publication Critical patent/WO2013159108A2/en
Publication of WO2013159108A3 publication Critical patent/WO2013159108A3/en

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/111General methods applicable to biologically active non-coding nucleic acids
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y301/00Hydrolases acting on ester bonds (3.1)
    • C12Y301/03Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03016Phosphoprotein phosphatase (3.1.3.16), i.e. calcineurin
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y301/00Hydrolases acting on ester bonds (3.1)
    • C12Y301/03Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03048Protein-tyrosine-phosphatase (3.1.3.48)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y301/00Hydrolases acting on ester bonds (3.1)
    • C12Y301/03Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03067Phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase (3.1.3.67)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/10Type of nucleic acid
    • C12N2310/11Antisense
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/32Chemical structure of the sugar
    • C12N2310/3212'-O-R Modification
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/32Chemical structure of the sugar
    • C12N2310/323Chemical structure of the sugar modified ring structure
    • C12N2310/3231Chemical structure of the sugar modified ring structure having an additional ring, e.g. LNA, ENA
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/34Spatial arrangement of the modifications
    • C12N2310/341Gapmers, i.e. of the type ===---===
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/34Spatial arrangement of the modifications
    • C12N2310/343Spatial arrangement of the modifications having patterns, e.g. ==--==--==--
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/35Nature of the modification
    • C12N2310/352Nature of the modification linked to the nucleic acid via a carbon atom
    • C12N2310/3525MOE, methoxyethoxy
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2320/00Applications; Uses
    • C12N2320/50Methods for regulating/modulating their activity
    • C12N2320/51Methods for regulating/modulating their activity modulating the chemical stability, e.g. nuclease-resistance

Abstract

The present invention provides oligomeric compounds. Certain such oligomeric compounds are useful for hybridizing to a complementary nucleic acid, including but not limited, to nucleic acids in a cell. In certain embodiments, hybridization results in modulation of the amount activity or expression of the target nucleic acid in a cell.

Description

OLIGOMERIC COMPOUNDS COMPRISING BICYCLIC NUCLEOTIDES AND USES THEREOF

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled CORE0104USLSEQ.txt, created April 20, 2012, which is 4 Kb in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

Antisense compounds have been used to modulate target nucleic acids. Antisense compounds comprising a variety of chemical modifications and motifs have been reported. In certain instances, such compounds are useful as research tools, diagnostic reagents, and as therapeutic agents. In certain instances antisense compounds have been shown to modulate protein expression by binding to a target messenger RNA (mRNA) encoding the protein. In certain instances, such binding of an antisense compound to its target mRNA results in cleavage of the mRNA. Antisense compounds that modulate processing of a pre -mRNA have also been reported. Such antisense compounds alter splicing, interfere with polyadenlyation or prevent formation of the 5 '-cap of a pre-mRNA.

Certain antisense compounds have been described previously. See for example United States Patent No. 7,399,845 and published International Patent Application No. WO 2008/049085, which are hereby incorporated by reference herein in their entirety.

SUMMARY OF THE INVENTION

In certain embodiments, the present invention provides compounds comprising oligonucleotides. In certain embodiments, such oligonucleotides comprise a gapmer region. In certain embodiments, such oligonucleotides consist of a gapmer region.

The present disclosure provides the following non- limiting numbered embodiments:

Embodiment 1 : A compound comprising:

a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide comprises:

a 5 '-wing consisting of 2 to 5 linked nucleosides;

a 3 '-wing consisting of 2 to 5 linked nucleosides; and

a gap between the 5'-wing and the 3'-wing consisting of 6 to 14 linked 2'-deoxynucleosides; and wherein at least one of the 5'-wing and the 3'-wing comprises at least one bicyclic nucleoside; at least one of the 5 '-wing and the 3 '-wing comprises at least one 2 '-substituted nucleoside; and

wherein the nucleobase sequence of the modified oligonucleotide is complementary to the nucleobase

sequence of a target nucleic acid.

Embodiment 2: The compound of embodiment 1, wherein one of the 5 '-wing or the 3 '-wing comprises at least one 2'-deoxynucleoside.

Embodiment 3: The compound of embodiments 1-2, wherein each of the 5 '-wing and the 3 '-wing comprises at least one 2'-deoxynucleoside.

Embodiment 4: The compound of embodiments 1-3, wherein the 3 '-wing comprises at least one 2'- deoxynucleoside.

Embodiment 5: The compound of embodiments 1-4, wherein the 5 '-wing comprises at least one 2'- deoxynucleoside.

Embodiment 6: The compound of any of embodiments 1-5, wherein the 5'-wing comprises at least one bicyclic nucleoside.

Embodiment 7: The compound of any of embodiments 1-6, wherein the 3 '-wing comprises at least one bicyclic nucleoside.

Embodiment 8: The compound of any of embodiments 1-7, wherein the 5'-wing comprises at least one 2'- substituted nucleoside.

Embodiment 9: The compound of any of embodiments 1-8, wherein the 3 '-wing comprises at least one 2'- substituted nucleoside.

Embodiment 10: A compound comprising:

a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide comprises:

a 5 '-wing consisting of 2 to 5 linked nucleosides;

a 3 '-wing of 2 to 5 linked nucleosides; and a gap between the 5' wing and the 3' wing consisting of 6 to 14 linked 2'-deoxynucleosides; and wherein at least one of the 5 '-wing and the 3 '-wing comprises at least one constrained ethyl nucleoside; and at least one of the 5 '-wing and the 3 '-wing comprises at least one 2 '-substituted nucleoside; and wherein the nucleobase sequence of the modified oligonucleotide is complementary to the nucleobase sequence of a target nucleic acid.

Embodiment 11 : The compound of embodiments 1-10, wherein and at least one of the 5 '-wing and the

3 '-wing comprises at least one 2'-deoxynucleoside.

Embodiment 12: The compound of embodiments 1-1 1, wherein at least one of the 5 '-wing and the 3'- wing comprises both at least one constrained ethyl nucleoside and at least one 2 '-substituted nucleoside.

Embodiment 13: The compound of embodiments 1-12, wherein the 5 '-wing comprises at least one constrained ethyl nucleoside.

Embodiment 14: The compound of any of embodiments 10-13, wherein the 3 '-wing comprises at least one constrained ethyl nucleoside.

Embodiment 15: The compound of any of embodiments 10-14, wherein the 5 '-wing comprises at least one 2 '-substituted nucleoside.

Embodiment 16: The compound of any of embodiments 10-15, wherein the 3'-wing comprises at least one 2 '-substituted nucleoside.

Embodiment 17: The compound of any of embodiments 1- 17, wherein the modified oligonucleotide has a sugar motif described by Formula I as follows:

(A)m-(B)n-(J)p-(B)r-(J)t-(D)g-(J)v-(B)w-(J)x-(B)y-(A)z

wherein:

each A is independently a 2 '-substituted nucleoside;

each B is independently a bicyclic nucleoside;

each J is independently either a 2 '-substituted nucleoside or a 2'-deoxynucleoside;

each D is a 2'-deoxynucleoside;

m is 0-4; n is 0-2; p is 0-2; r is 0-2; t is 0-2; v is 0-2; w is 0-4; x is 0-2; y is 0-2; z is 0-4; and g is 6- 14;

provided that: at least one of m, n, and r is other than 0;

at least one of w and y is other than 0;

the sum of m, n, p, r, and t is from 2 to 5; and

the sum of v, w, x, y, and z is from 2 to 5.

Embodiment 18: A compound comprising:

a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide has a sugar motif described by Formula I as follows:

(A)m-(B)n-(J)p-(B)r-(J)r(D)g-(J)v-(B)w-(J)x-(B)y-(A)z

wherein:

each A is independently a 2 '-substituted nucleoside;

each B is independently a bicyclic nucleoside;

each J is independently either a 2 '-substituted nucleoside or a 2'-deoxynucleoside;

each D is a 2'-deoxynucleoside;

m is 0-4; n is 0-2; p is 0-2; r is 0-2; t is 0-2; v is 0-2; w is 0-4; x is 0-2; y is 0-2; z is 0-4; and g is 6- 14;

provided that:

at least one of m, n, and r is other than 0;

at least one of w and y is other than 0;

the sum of m, n, p, r, and t is from 2 to 5; and

the sum of v, w, x, y, and z is from 2 to 5.

Embodiment 19: The compound of embodiment 17 or 18, wherein at least one bicyclic nucleoside is a constrained ethyl nucleoside.

Embodiment 20: The compound of embodiment 17 or 18, wherein each bicyclic nucleoside is a

constrained ethyl nucleoside.

Embodiment 21 : The compound of any of embodiments 17- 19, wherein at least one bicyclic

nucleoside is an LNA nucleoside.

Embodiment 22: The compound of embodiment 17 or 18, wherein each bicyclic nucleoside is an LNA nucleoside. Embodiment 23: The compound of any of embodiments 1-22, wherein the 2'-substituent of the at least one 2 '-substituted nucleoside is selected from among: OCH3, F, OCH2F, OCHF2, OCF3, OCH2CH3, 0(CH2)2F, OCH2CHF2, OCH2CF3, OCH2-CH=CH2, 0(CH2)2-OCH3, 0(CH2)2-SCH3, 0(CH2)2-OCF3, 0(CH2)3-N(R4)(R5), 0(CH2)2-ON(R4)(R5), 0(CH2)2-0(CH2)2-N(R4)(R5), OCH2C(=0)-N(R4)(R5), OCH2C(=0)-N(R6)-(CH2)2-N(R4)(R5) and 0(CH2)2-N(R6)-C(=NR7)[N(R4)(R5)] wherein R4, R5, Re and R7 are each, independently, H or Ci-C6 alkyl.

Embodiment 24: The compound of embodiment 23, wherein the 2'-substituent of the at least one 2'- substituted nucleoside of is selected from among: OCH3, F, and 0(CH2)2-OCH3.

Embodiment 25: The compound of embodiment 24, wherein the 2'-substituent of the at least one 2'- substituted nucleoside is 0(CH2)2-OCH3.

Embodiment 26: The compound of any of embodiments 1-22, wherein the 2'-substituent of each 2'- substituted nucleoside is selected from among: OCH3, F, OCH2F, OCHF2, OCF3, OCH2CH3, 0(CH2)2F, OCH2CHF2, OCH2CF3, OCH2-CH=CH2, 0(CH2)2-OCH3, 0(CH2)2-SCH3, 0(CH2)2-OCF3, 0(CH2)3- N(R4)(R5), 0(CH2)2-ON(R4)(R5), 0(CH2)2-0(CH2)2-N(R4)(R5), OCH2C(=0)-N(R4)(R5), OCH2C(=0)- N(R6)-(CH2)2-N(R4)(R5) and 0(CH2)2-N(R6)-C(=NR7)[N(R4)(R5)] wherein R4, R5, R6 and R7 are each, independently, H or C1-C6 alkyl.

Embodiment 27: The compound of embodiment 26, wherein the 2'-substituent of each 2 '-substituted nucleoside of is selected from among: OCH3, F, and 0(CH2)2-OCH3.

Embodiment 28: The compound of embodiment 27, wherein the 2'-substituent of each 2 '-substituted nucleoside is 0(CH2)2-OCH3.

Embodiment 29: The compound of any of embodiments 1-28, wherein the 5'-wing does not comprise a bicyclic nucleotide.

Embodiment 30: The compound of any of embodiments 1-29, wherein the 3 '-wing does not comprise a bicyclic nucleotide.

Embodiment 31 : The compound of any of embodiments 1-30, wherein the target nucleic acid is not a

Huntingtin gene transcript. Embodiment 32: The compound of any of embodiments 1-31, wherein the modified oligonucleotide has a base sequence other than:

GTGCTACCCAACCTTTCTG (SEQ ID NO: 1); CACAGTGCTACCCAACCTT (SEQ ID NO: 2);

CAGTGCTACCCAACC (SEQ ID NO: 3); ATATCACAGTGCTACCCAA (SEQ ID NO: 4);

GATGCTGACTTGGGCCATT(SEQ ID NO: 5); GGGATGCTGACTTGG (SEQ ID NO: 6);

TGCCAAGGGATGCTGACTT(SEQ ID NO: 7); AATTGTCATCACCAGAAAA (SEQ ID NO: 8);

TAAATTGTCATCACC (SEQ ID NO: 9); ACAGTAGATGAGGGAGCAG (SEQ ID NO: 10);

ACACAGTAGATGAGG (SEQ ID NO: 1 1); AAGTGCACACAGTAGATGA (SEQ ID NO: 12);

AGCTGCAACCTGGCAACAA (SEQ ID NO: 13); GCAGCTGCAACCTGG (SEQ ID NO: 14); or GCAAGAGCAGCTGCAACCT (SEQ ID NO: 15).

Embodiment 33: The compound of any of embodiments 1-31, wherein the oligonucleotide has a sugar motif other than:

E-K-K-(D)9-K-K-E;

E-E-E-E-K-(D)9-E-E-E-E-E;

E-K-K-K-(D)9-K-K-K-E;

K-E-E-K-(D)9-K-E-E-K;

K-D-D-K-(D)9-K-D-D-K;

K-E-K-E-K-(D)9-K-E-K-E-K;

K-D-K-D-K-(D)9-K-D-K-D-K;

E-K-E-K-(D)9-K-E-K-E;

E-E-E-E-E-K-(D)8-E-E-E-E-E; or

E-K-E-K-E-(D)9-E-K-E-K-E; wherein

K is a constrained ethyl nucleoside, E is a 2'-MOE substituted nucleoside, and D is a 2'- deoxynucleoside.

Embodiment 34: The compound of any of embodiments 1-30, wherein the 5'-wing consists of 2 linked nucleosides.

Embodiment 35: The compound of any of embodiments 1-30, wherein the 5'-wing consists of 3 linked nucleosides.

Embodiment 36: The compound of any of embodiments 1-30, wherein the 5'-wing consists of 4 linked nucleosides. Embodiment 37: The compound of any of embodiments 1-30, wherein the 5'-wing consists of 5 linked nucleosides.

Embodiment 38: The compound of any of embodiments 1-34, wherein the 3'-wing consists of 2 linked nucleosides.

Embodiment 39: The compound of any of embodiments 1-34, wherein the 3 '-wing consists of 3 linked nucleosides.

Embodiment 40: The compound of any of embodiments 1-34, wherein the 3 '-wing consists of 4 linked nucleosides.

Embodiment 41 : The compound of any of embodiments 1-34, wherein the 3'-wing consists of 5 linked nucleosides.

Embodiment 42: The compound of any of embodiments 1-38, wherein the gap consists of 6 linked 2'- deoxynucleosides.

Embodiment 43: The compound of any of embodiments 1-38, wherein the gap consists of 7 linked 2'- deoxynucleosides.

Embodiment 44: The compound of any of embodiments 1-38, wherein the gap consists of 8 linked 2'- deoxynucleosides.

Embodiment 45: The compound of any of embodiments 1-38, wherein the gap consists of 9 linked 2'- deoxynucleosides.

Embodiment 46: The compound of any of embodiments 1-38, wherein the gap consists of 10 linked 2'-deoxynucleosides.

Embodiment 47: The compound of any of embodiments 1-38, wherein the gap consists of 11 linked 2'-deoxynucleosides.

Embodiment 48: The compound of any of embodiments 1-38, wherein the gap consists of 12 linked 2'-deoxynucleosides. Embodiment 49: The compound of any of embodiments 1-38, wherein the gap consists of 13 linked

2'-deoxynucleosides.

Embodiment 50: The compound of any of embodiments 1-38, wherein the gap consists of 14 linked

2'-deoxynucleosides.

Embodiment 51 : The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

10 linked nucleosides.

Embodiment 52: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

1 1 linked nucleosides.

Embodiment 53: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

12 linked nucleosides.

Embodiment 54: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

13 linked nucleosides.

Embodiment 55: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

14 linked nucleosides.

Embodiment 56: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

15 linked nucleosides.

Embodiment 57: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

16 linked nucleosides.

Embodiment 58: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

17 linked nucleosides.

Embodiment 59: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

18 linked nucleosides.

Embodiment 60: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

19 linked nucleosides. Embodiment 61 : The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

20 linked nucleosides.

Embodiment 62: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

21 linked nucleosides.

Embodiment 63: The compound of any of embodiments 1-50, wherein the oligonucleotide consists of

22 linked nucleosides.

Embodiment 64: The compound of any of embodiments 1-30, wherein the gapmer motif is selected from among: 2-10-2, 2-10-3, 2-10-4, 2-10-5, 3-10-2, 3-10-3, 3-10-4, 3- 10-5, 4-10-2, 4-10-3, 4-10-4, 4- 10-5, 5-10-2, 5- 10-3, 5-10-4, 5- 10-5, 2-9-2, 2-9-3, 2-9-4, 2-9-5, 3-9-2, 3-9-3, 3-9-4, 3-9-5, 4-9-2, 4-9-3, 4-9-4, 4-9-5, 5-9-2, 5-9-3, 5-9-4, 5-9-5, 2-8-2, 2-8-3, 2-8-4, 2-8-5, 3-8-2, 3-8-3, 3-8-4, 3-8-5, 4-8-2, 4-8- 3, 4-8-4, 4-8-5, 5-8-2, 5-8-3, 5-8-4, and 5-8-5.

Embodiment 65: A compound comprising a modified oligonucleotide having a sugar motif selected from among sugar motifs 1-278 as shown in Table 4.

Embodiment 66: The compound of any of embodiments 1-65, wherein the 5'-wing has a motif

selected from among the 5 '-wing motifs as shown in Tables 1-3.

Embodiment 67: The compound of any of embodiments 1-66, wherein the 3 '-wing has a motif

selected from among the 3 '-wing motifs as shown in Tables 4-6.

Embodiment 68: The compound of any of embodiments 66-67, wherein each A, each B, and each C are independently selected from among: HNA and F-HNA.

Embodiment 69: The compound of any of embodiments 1-68, wherein the 5'-wing comprises at least one F-HNA.

Embodiment 70: The compound of any of embodiments 1-69, wherein the 3 '-wing comprises at least one F-HNA.

Embodiment 71 : The compound of any of embodiments 1-68, wherein the 5'-wing comprises at least one modified nucleobase. Embodiment 72: The compound of any of embodiments 1-69, wherein the 3 '-wing comprises at least one modified nucleobase.

Embodiment 73 : The compound of embodiment 72, wherein the modified nucleobase is 2-thio- thymidine.

Embodiment 74: The compound of any of embodiments 1-73, wherein the 5'-wing has a motif

selected from among the 5 '-wing motifs as shown in Tables 1-3 and the 3 '-wing has a motif selected from among the 3 '-wing motifs as shown in Tables 4-6.

Embodiment 75: The compound of any of embodiments 1-74, wherein the 5'-wing has an ABABA motif, wherein each A is a modified nucleoside and each B comprises a 2'-deoxynucleoside.

Embodiment 76: The compound of embodiment 75, wherein the modified nucleoside is a bicyclic nucleoside.

Embodiment 77: The compound of embodiment 76, wherein the bicyclic nucleoside is cEt.

Embodiment 78: The compound of embodiment 76, wherein the bicyclic nucleoside is LNA.

Embodiment 79: The compound of any of embodiments 75-78 wherein the 3 '-wing has a motif selected from among: AA, AB, AC, BA, BB, BC, CA, CB, and CC.

Embodiment 80: The compound of embodiment 79, wherein the 3 '-wing has an AA motif.

Embodiment 81 : The compound of embodiment 80, wherein A is a 2 '-substituted nucleoside.

Embodiment 82: The compound of embodiment 80, wherein the 2 '-substituted nucleoside is selected from among: OCH3, F, OCH2F, OCHF2, OCF3, OCH2CH3, 0(CH2)2F, OCH2CHF2, OCH2CF3, OCH2- CH=CH2, 0(CH2)2-OCH3, 0(CH2)2-SCH3, 0(CH2)2-OCF3, 0(CH2)3-N(R4)(R5), 0(CH2)2-ON(R4)(R5), 0(CH2)2-0(CH2)2-N(R4)(R5), OCH2C(=0)-N(R4)(R5), OCH2C(=0)-N(R6)-(CH2)2-N(R4)(R5) and 0(CH2)2-N(R6)-C(=NR7)[N(R4)(R5)] wherein R4, R5, R6 and R7 are each, independently, H or C C6 alkyl.

Embodiment 83: The compound of embodiment 82, wherein the 2'-substituent of each 2 '-substituted nucleoside of is selected from among: OCH3, F, and 0(CH2)2-OCH3. Embodiment 84: The compound of embodiment 83, wherein the 2'-substituent of each 2 '-substituted nucleoside is 0(CH2)2-OCH3.

Embodiment 85: The compound of any of embodiments 76-84 wherein the gap between the 5 '-wing and the 3'-wing consists of 6 to 1 1 linked 2'-deoxynucleosides.

Embodiment 86: The compound of any of embodiments 76-84 wherein the gap between the 5'-wing and the 3'-wing consists of 7 to 10 linked 2'-deoxynucleosides.

Embodiment 87: The compound of any of embodiments 76-84 wherein the gap between the 5'-wing and the 3'-wing consists of 10 linked 2'-deoxynucleosides.

Embodiment 88: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D- - (D)6-E-E.

Embodiment 89: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D-K- (D)7-E-E.

Embodiment 90: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D- - (D)g-E-E.

Embodiment 91 : The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D-K-

(D)9-E-E.

Embodiment 92: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D- - (D)io-E-E.

Embodiment 93 : The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D-K- (D)i i-E-E.

Embodiment 94: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D- - (D)i2-E-E.

Embodiment 95: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D-K- (D)B-E-E. Embodiment 96: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D-K-

(D)i4-E-E.

Embodiment 97: The compound of any of embodiments 75-87 having the sugar motif: K-D-K-D-K-

(D)15-E-E.

Embodiment 98: The compound of any of embodiments 1-97, wherein the 5'-wing has a BDBDB motif, wherein each B is a bicyclic nucleoside and each D comprises a 2'-deoxynucleoside.

Embodiment 99: The compound of any of embodiments 1-97, wherein the 5'-wing has a BDBDB-

(D)6_i5-AA motif, wherein each B is a bicyclic nucleoside and each D comprises a 2'-deoxynucleoside.

Embodiment 100: The compound of any of embodiments 98-99, wherein B is selected from among:

BNA, LNA, a-L-LNA, ENA and 2'-thio LNA.

Embodiment 101 : The compound of embodiment 100, wherein B comprises BNA.

Embodiment 102: The compound of embodiment 100, wherein B comprises LNA.

Embodiment 103: The compound of embodiment 100, wherein B comprises a-L-LNA.

Embodiment 104: The compound of embodiment 100, wherein B comprises ENA.

Embodiment 105: The compound of embodiment 100, wherein B comprises 2'-thio LNA.

Embodiment 106: The compound of any of embodiments 100 to 105, wherein A comprises a

2'substituted nucleoside.

Embodiment 107: The compound of cliam 106, wherein the 2' substituted nucleoside comprises MOE.

Embodiment 108: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-B-B-(D)8-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside Embodiment 109: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-B-B-(D)9-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 110: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-B-B-(D)10-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 11 1 : The compound of any of embodiments 1 -2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-B-(D)8-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 112: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-B-(D)9-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 113: The compound of any of embodiments 1 -2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-B-(D)i0-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 114: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-B-(D)8-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 115: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-B-(D)9-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside Embodiment 116: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-B-(D)i0-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 117: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-A-(D)8-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 118: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-A-(D)9-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 119: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-A-(D)i0-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 120: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-(D)8-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 121 : The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-(D)9-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 122: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-(D)i0-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside Embodiment 123: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-(D)8-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 124: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-(D)9-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 125: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-(D)i0-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 126: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-(D)8-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 127: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-(D)9-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 128: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-(D)i0-B-B-A, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 129: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-(D)8-B-B-B, wherein each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside Embodiment 130: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-(D)9-B-B-B, wherein each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 131 : The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-(D)10-B-B-B, wherein each B is an

independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 132: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-(D)8-B-B-B, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 133: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-(D)9-B-B-B, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 134: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-(D)i0-B-B-B, wherein each A is an

independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 135: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-D-B-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 136: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-D-B-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 137: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-D-B-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 138: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-D-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 139: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-D-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 140: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-D-A-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 141 : The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-B-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 142: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-B-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 143: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-B-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 144: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 145: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 146: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 147: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 148: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 149: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-(D)10-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 150: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-(D)8-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 151 : The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-(D)9-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 152: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-(D)10-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 153: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-D-B-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 154: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-D-B-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 155: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-D-B-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 156: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-(D)8-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 157: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-(D)9-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 158: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-(D)i0-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 159: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-B-(D)8-B-B-B, wherein each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 160: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-B-(D)9-B-B-B, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 161 : The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-B-B-B-(D)i0-B-B-B, wherein each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 162: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 163: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 164: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-A-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 165: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-A-(D)8-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside Embodiment 166: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-A-(D)9-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 167: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-A-(D)10-B-B-B, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 168: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-A-D-B-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 169: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-A-D-B-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 170: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-A-D-B-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 171 : The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-B-D-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 172: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-B-D-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside Embodiment 173: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-D-B-D-A-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 174: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-A-D-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 175: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-A-D-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 176: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-D-A-D-A-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 177: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-B-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 178: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-B-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 179: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-A-A-B-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside Embodiment 180: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-B-A-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 181 : The compound of any of embodiments 1 -2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-B-A-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 182: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: A-A-B-A-A-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 183: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-A-(D)8-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 184: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-A-(D)9-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 185: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: B-A-A-A-A-(D)i0-B-B-A, wherein each A is an independently selected 2 '-substituted nucleoside, each B is an independently selected bicyclic nucleoside, and each D is a 2'-deoxynucleoside

Embodiment 186: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-k-k-(D)9-e-k-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside. Embodiment 187: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-d-k-d-k-(D)i0-e-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 188: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-e-k-(D)10-k-e-k, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.

Embodiment 189: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-d-k-d-k-(D)8-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 190: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-d-k-d-k-(D)8-e-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 191 : The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-e-k-(D)8-e-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 192: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-k-(D)10-k-e-k-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.

Embodiment 193: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-(D)i0-k-k-k, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.

Embodiment 194: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-k-(D)8-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside. Embodiment 195: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-k-(D)7-k-k-e-e-e, wherein each k comprises bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 196: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-(D)9-k-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 197: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-k-(D)7-k-k-e-e-e, wherein each k comprises bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 198: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-k-(D)7-k-k-e-e-e, wherein each k comprises bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 199: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-e-k-k-(D)7-e-e-e-e, wherein each k comprises bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 200: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-k-e-k-(D)9-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 201 : The compound of any of embodiments 1 -2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-k-e-k-d-k-(D)7-e-e-e-e, wherein each k comprises bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside. Embodiment 202: The compound of any of embodiments 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-k-(D)7-k-k-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'- deoxynucleoside.

Embodiment 203: The compound of any of embodiments 186 to 202, wherein each k comprises a cEt nucleoside.

Embodiment 204: The compound of any of embodiments 186 to 202, wherein each k comprises an LNA nucleoside.

Embodiment 205: The compound of any of embodiments 186 to 203, wherein each e comprises a 2'- MOE modified nucleoside.

Embodiment 206: The compound of any of embodiments 186 to 203, wherein each e comprises a 2'- OMe modified nucleoside.

Embodiment 207: The compound of any of embodiments 186 to 202, wherein each k comprises a cEt nucleoside and each e comprises a 2'-MOE modified nucleoside.

Embodiment 208: The compound of any of embodiments 89-202, wherein at least one bicyclic

nucleoside is a constrained ethyl nucleoside.

Embodiment 209: The compound of any of embodiments 89-202, wherein each bicyclic nucleoside is a constrained ethyl nucleoside.

Embodiment 210: The compound of any of embodiments, 89-202, wherein at least one bicyclic

nucleoside is selected from among: BNA, LNA, a-L-LNA, ENA and 2'-thio LNA.

Embodiment 21 1 : The compound of any of embodiments, 89-202, wherein at least one bicyclic

nucleoside is an LNA nucleoside.

Embodiment 212: The compound of any of embodiments 89-202, wherein each bicyclic nucleoside is an LNA nucleoside. Embodiment 213: The compound of any of embodiments 89-202, wherein the 2'-substituent of the at least one 2 '-substituted nucleoside is selected from among: OCH3, F, OCH2F, OCHF2, OCF3, OCH2CH3, 0(CH2)2F, OCH2CHF2, OCH2CF3, OCH2-CH=CH2, 0(CH2)2-OCH3, 0(CH2)2-SCH3, 0(CH2)2-OCF3, 0(CH2)3-N(R4)(R5), 0(CH2)2-ON(R4)(R5), 0(CH2)2-0(CH2)2-N(R4)(R5), OCH2C(=0)-N(R4)(R5), OCH2C(=0)-N(R6)-(CH2)2-N(R4)(R5) and 0(CH2)2-N(R6)-C(=NR7)[N(R4)(R5)] wherein R4, R5, Re and R7 are each, independently, H or Ci-C6 alkyl.

Embodiment 214: The compound of embodiment 213, wherein the 2'-substituent of the at least one 2'- substituted nucleoside of is selected from among: OCH3, F, and 0(CH2)2-OCH3.

Embodiment 215: The compound of embodiment 214, wherein the 2'-substituent of the at least one 2'- substituted nucleoside is 0(CH2)2-OCH3.

Embodiment 216: The compound of any of embodiments 89-202, wherein the 2'-substituent of each 2'- substituted nucleoside is selected from among: OCH3, F, OCH2F, OCHF2, OCF3, OCH2CH3, 0(CH2)2F, OCH2CHF2, OCH2CF3, OCH2-CH=CH2, 0(CH2)2-OCH3, 0(CH2)2-SCH3, 0(CH2)2-OCF3, 0(CH2)3- N(R4)(R5), 0(CH2)2-ON(R4)(R5), 0(CH2)2-0(CH2)2-N(R4)(R5), OCH2C(=0)-N(R4)(R5), OCH2C(=0)- N(R6)-(CH2)2-N(R4)(R5) and 0(CH2)2-N(R6)-C(=NR7)[N(R4)(R5)] wherein R4, R5, R6 and R7 are each, independently, H or C1-C6 alkyl.

Embodiment 217: The compound of embodiment 216, wherein the 2'-substituent of each 2 '-substituted nucleoside of is selected from among: OCH3, F, and 0(CH2)2-OCH3.

Embodiment 218: The compound of embodiment 217, wherein the 2'-substituent of each 2 '-substituted nucleoside is 0(CH2)2-OCH3.

Embodiment 219: The compound of any of embodiments 1-218, wherein the oligonucleotide comprises at least one modified internucleoside linkage.

Embodiment 220: The compound of embodiment 219, wherein each internucleoside linkage is a

modified internucleoside linkage.

Embodiment 221 : The compound of embodiment 219 or 220, wherein the modified internucleoside linkage is a phosphorothioate linkage. Embodiment 222: The compound of embodiment 219 or 220, wherein the modified internucleoside linkage is a methylphosphonate.

Embodiment 223 : The compound of any of embodiments 1 -222 comprising a conjugate.

Embodiment 224: The compound of any of embodiments 1-223 comprising at least one 5-methyl cytosine nucleobase.

Embodiment 225: The compound of any of embodiments 1-224 comprising at least one modified nucleobase.

Embodiment 226: The compound of any of embodiments 1-225, wherein the compound is an antisense compound.

Embodiment 227: The compound of embodiment 226, wherein the compound is capable of inhibiting expression of the target nucleic acid in a cell.

Embodiment 228: The compound of embodiment 227, wherein the compound is capable of inhibiting expression of the target nucleic acid in a cell by at least 50%.

Embodiment 229: The compound of embodiment 227, wherein the compound is capable of inhibiting expression of the target nucleic acid in a cell by at least 80%.

Embodiment 230: The compound of any of embodiments 227-229, wherein the cell is in an animal.

Embodiment 231 : The compound of embodiment 230, wherein the animal is a human.

Embodiment 232: The compound of any of embodiments 1 to 231, wherein bicyclic nucleoside is selected from among: BNA, LNA, a-L-LNA, ENA and 2'-thio LNA.

Embodiment 233: A compound of any of embodiments 1-232, comprising not more than 6 bicyclic nucleosides.

Embodiment 234: A compound of any of embodiments 1-232, comprising not more than 5 bicyclic nucleosides. Embodiment 235: A compound of any of embodiments 1-232, comprising not more than 4 bicyclic nucleosides.

Embodiment 236: A compound of any of embodiments 1-232, comprising not more than 3 bicyclic nucleosides.

Embodiment 237: A compound of any of embodiments 1-232, comprising not more than 2 bicyclic nucleosides.

Embodiment 238: A compound of any of embodiments 1-232, comprising not more than 1 bicyclic nucleoside.

Embodiment 239: The compound of any of embodiments 233-238, wherein the bicyclic nucleoside comprises cEt.

Embodiment 240: The compound of any of embodiments 233-238, wherein the bicyclic nucleoside comprises LNA.

Embodiment 241 : A pharmaceutical composition comprising the compound according to any of embodiments 1 -240 and a pharmaceutically acceptable diluent.

Embodiment 242: A method of modulating expression of a target nucleic acid in a cell comprising contacting the cell with a compound according to any of embodiments 1-240.

Embodiment 243 : A method of modulating expression of a target nucleic acid in an animal comprising administering to the animal the pharmaceutical composition according to embodiment 242.

Embodiment 244: A method of manufacturing a compound according to any of embodiments 1-241 comprising forming chemical bonds.

Embodiment 245: The method of embodiment 244, wherein said chemical bonds are internucleoside linkages.

Embodiment 246: The method embodiment 244 or 245, wherein the method is performed under conditions suitable for the preparation of products for administration to humans. Embodiment 247: A method of manufacturing the pharmaceutical composition according to embodiment 246 comprising combining the compound according to any of embodiments 1 -241 and the pharmaceutically acceptable diluent.

Embodiment 248: The method embodiment 247, wherein the method is performed under conditions suitable for the preparation of products for administration to humans.

Embodiment 249: A compound comprising a modified oligonucleotide having a sugar motif selected from among sugar motifs 279-615 as shown in Table 4.

Embodiment 250: A compound comprising:

a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide comprises:

a 5 '-wing consisting of 2 to 5 linked nucleosides;

a 3 '-wing consisting of 2 to 5 linked nucleosides; and

a gap between the 5'-wing and the 3'-wing consisting of 6 to 14 linked 2'-deoxynucleosides; and wherein the 5 '-wing has a sugar modification motif selected from among the motifs in Table 1.

Embodiment 251 : A compound comprising:

a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide comprises:

a 5 '-wing consisting of 2 to 5 linked nucleosides;

a 3 '-wing consisting of 2 to 5 linked nucleosides; and

a gap between the 5'-wing and the 3'-wing consisting of 6 to 14 linked 2'-deoxynucleosides; and wherein the 3 '-wing has a sugar modification motif selected from among the motifs in Table 2.

Embodiment 252: A compound comprising:

a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide comprises:

a 5 '-wing consisting of 2 to 5 linked nucleosides;

a 3 '-wing consisting of 2 to 5 linked nucleosides; and a gap between the 5'-wing and the 3'-wing consisting of 6 to 14 linked 2'-deoxynucleosides; and wherein the 5 '-wing has a sugar modification motif selected from among the motifs in Table 1 and the 3'- wing has a sugar modification motif selected from among the motifs in Table 2.

Embodiment 253: A compound of any of embodiments 1-16, wherein the modified oligonucleotide has a sugar motif described by Formula II as follows:

(J)m-(B)n-(J)p-(B)r-(A)t-(D)g-(A)v-(B)w-(J)x-(B)y-(J)z

wherein:

each A is independently a 2 '-substituted nucleoside;

each B is independently a bicyclic nucleoside;

each J is independently either a 2 '-substituted nucleoside or a 2'-deoxynucleoside;

each D is a 2'-deoxynucleoside;

m is 0-4; n is 0-2; p is 0-2; r is 0-2; t is 0-2; v is 0-2; w is 0-4; x is 0-2; y is 0-2; z is 0-4; g is 6-14; provided that:

at least one of m, n, and r is other than 0;

at least one of w and y is other than 0;

the sum of m, n, p, r, and t is from 1 to 5; and

the sum of v, w, x, y, and z is from 1 to 5.

Embodiment 254: A compound comprising:

a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide has a sugar motif described by Formula II as follows:

(J)m-(B)n-(J)p-(B)r-(A)t-(D)g-(A)v-(B)w-(J)x-(B)y-(J)z

wherein:

each A is independently a 2 '-substituted nucleoside;

each B is independently a bicyclic nucleoside;

each J is independently either a 2 '-substituted nucleoside or a 2'-deoxynucleoside;

each D is a 2'-deoxynucleoside;

m is 0-4; n is 0-2; p is 0-2; r is 0-2; t is 0-2; v is 0-2; w is 0-4; x is 0-2; y is 0-2; z is 0-4; g is 6-14; provided that:

at least one of m, n, and r is other than 0;

at least one of w and y is other than 0;

the sum of m, n, p, r, and t is from 1 to 5; and

the sum of v, w, x, y, and z is from 1 to 5. Embodiment 255: The compound of embodiment 253 or 254, wherein at least one bicyclic nucleoside is a constrained ethyl nucleoside.

Embodiment 256: The compound of embodiment 255, wherein each bicyclic nucleoside is a

constrained ethyl nucleoside.

Embodiment 257: The compound of any of embodiments 253 -254, wherein at least one bicyclic

nucleoside is an LNA nucleoside.

Embodiment 258: The compound of embodiments 250-254, wherein each bicyclic nucleoside is an LNA nucleoside.

Embodiment 259: A method of treating a disease or condition.

Embodiment 260: Use of a compound of any of embodiments 1 to 259 for the preparation of a

medicament for the treatment of a disease or condition.

Embodiment 261 : The use of embodiment 260, wherein the disease or condition is associated with a virus.

In certain embodiments, including but not limited to any of the above numbered embodiments, compounds including oligonucleotides described herein are capable of modulating expression of a target mRNA. In certain embodiments, the target mRNA is associated with a disease or disorder, or encodes a protein that is associated with a disease or disorder. In certain embodiments, the compounds or

oligonucleotides provided herein modulate the expression of function of such mRNA to alleviate one or more symptom of the disease or disorder.

In certain embodiments, compounds including oligonucleotides describe herein are useful in vitro. In certain embodiments such compounds are used in diagnostics and/or for target validation experiments.

DETAILED DESCRIPTION OF THE INVENTION

Unless specific definitions are provided, the nomenclature used in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical synthesis, and chemical analysis. Certain such techniques and procedures may be found for example in "Carbohydrate Modifications in Antisense Research" Edited by Sangvi and Cook, American Chemical Society , Washington D.C., 1994; "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, Pa., 21st edition, 2005; and "Antisense Drug Technology, Principles, Strategies, and Applications" Edited by Stanley T. Crooke, CRC Press, Boca Raton, Florida; and Sambrook et al., "Molecular Cloning, A laboratory Manual," 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, which are hereby incorporated by reference for any purpose. Where permitted, all patents, applications, published applications and other publications and other data referred to throughout in the disclosure are incorporated by reference herein in their entirety.

Unless otherwise indicated, the following terms have the following meanings:

As used herein, "nucleoside" means a compound comprising a nucleobase moiety and a sugar moiety. Nucleosides include, but are not limited to, naturally occurring nucleosides (as found in DNA and RNA) and modified nucleosides. Nucleosides may be linked to a phosphate moiety.

As used herein, "chemical modification" means a chemical difference in a compound when compared to a naturally occurring counterpart. In reference to an oligonucleotide, chemical modification does not include differences only in nucleobase sequence. Chemical modifications of oligonucleotides include nucleoside modifications (including sugar moiety modifications and nucleobase modifications) and internucleoside linkage modifications.

As used herein, "furanosyl" means a structure comprising a 5-membered ring comprising four carbon atoms and one oxygen atom.

As used herein, "naturally occurring sugar moiety" means a ribofuranosyl as found in naturally occurring RNA or a deoxyribofuranosyl as found in naturally occurring DNA.

As used herein, "sugar moiety" means a naturally occurring sugar moiety or a modified sugar moiety of a nucleoside.

As used herein, "modified sugar moiety" means a substituted sugar moiety or a sugar surrogate.

As used herein, "substituted sugar moiety" means a furanosyl that is not a naturally occurring sugar moiety. Substituted sugar moieties include, but are not limited to furanosyls comprising substituents at the 2 '-position, the 3 '-position, the 5 '-position and/or the 4 '-position. Certain substituted sugar moieties are bicyclic sugar moieties.

As used herein, "2 '-substituted sugar moiety" means a furanosyl comprising a substituent at the 2'- position other than H or OH. Unless otherwise indicated, a 2 '-substituted sugar moiety is not a bicyclic sugar moiety (i.e., the 2 '-substituent of a 2 '-substituted sugar moiety does not form a bridge to another atom of the furanosyl ring.

As used herein, "MOE" means -OCH2CH2OCH3.

As used herein the term "sugar surrogate" means a structure that does not comprise a furanosyl and that is capable of replacing the naturally occurring sugar moiety of a nucleoside, such that the resulting nucleoside is capable of (1) incorporation into an oligonucleotide and (2) hybridization to a complementary nucleoside. Such structures include rings comprising a different number of atoms than furanosyl (e.g., 4, 6, or 7-membered rings); replacement of the oxygen of a furanosyl with a non-oxygen atom (e.g., carbon, sulfur, or nitrogen); or both a change in the number of atoms and a replacement of the oxygen. Such structures may also comprise substitutions corresponding to those described for substituted sugar moieties (e.g., 6-membered carbocyclic bicyclic sugar surrogates optionally comprising additional substituents). Sugar surrogates also include more complex sugar replacements (e.g., the non-ring systems of peptide nucleic acid). Sugar surrogates include without limitation morpholinos, cyclohexenyls and cyclohexitols.

As used herein, "bicyclic sugar moiety" means a modified sugar moiety comprising a 4 to 7 membered ring (including but not limited to a furanosyl) comprising a bridge connecting two atoms of the 4 to 7 membered ring to form a second ring, resulting in a bicyclic structure. In certain embodiments, the 4 to 7 membered ring is a sugar ring. In certain embodiments the 4 to 7 membered ring is a furanosyl. In certain such embodiments, the bridge connects the 2 '-carbon and the 4 '-carbon of the furanosyl.

As used herein, "nucleotide" means a nucleoside further comprising a phosphate linking group. As used herein, "linked nucleosides" may or may not be linked by phosphate linkages and thus includes, but is not limited to "linked nucleotides." As used herein, "linked nucleosides" are nucleosides that are connected in a continuous sequence (i.e. no additional nucleosides are present between those that are linked).

As used herein, "nucleobase" means a group of atoms that can be linked to a sugar moiety to create a nucleoside that is capable of incorporation into an oligonucleotide, and wherein the group of atoms is capable of bonding with a complementary naturally occurring nucleobase of another oligonucleotide or nucleic acid. Nucleobases may be naturally occurring or may be modified.

As used herein, "heterocyclic base" or "heterocyclic nucleobase" means a nucleobase comprising a heterocyclic structure.

As used herein the terms, "unmodified nucleobase" or "naturally occurring nucleobase" means the naturally occurring heterocyclic nucleobases of RNA or DNA: the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) (including 5-methyl C), and uracil (U).

As used herein, "modified nucleobase" means any nucleobase that is not a naturally occurring nucleobase.

As used herein, "modified nucleoside" means a nucleoside comprising at least one chemical modification compared to naturally occurring RNA or DNA nucleosides. Modified nucleosides comprise a modified sugar moiety and/or a modified nucleobase.

As used herein, "bicyclic nucleoside" or "BNA" means a nucleoside comprising a bicyclic sugar moiety.

As used herein, "constrained ethyl nucleoside" or "cEt" means a nucleoside comprising a bicyclic sugar moiety comprising a 4'-CH(CH3)-0-2'bridge.

As used herein, "locked nucleic acid nucleoside" or "LNA" means a nucleoside comprising a bicyclic sugar moiety comprising a 4'-CH2-0-2'bridge.

As used herein, "2 '-substituted nucleoside" means a nucleoside comprising a substituent at the 2'- position other than H or OH. Unless otherwise indicated, a 2 '-substituted nucleoside is not a bicyclic nucleoside. As used herein, "2'-deoxynucleoside" means a nucleoside comprising 2'-H furanosyl sugar moiety, as found in naturally occurring deoxyribonucleosides (DNA). In certain embodiments, a 2'-deoxynucleoside may comprise a modified nucleobase or may comprise an RNA nucleobase (e.g., uracil).

As used herein, "oligonucleotide" means a compound comprising a plurality of linked nucleosides. In certain embodiments, an oligonucleotide comprises one or more unmodified ribonucleosides (RNA) and/or unmodified deoxyribonucleosides (DNA) and/or one or more modified nucleosides.

As used herein "oligonucleoside" means an oligonucleotide in which none of the internucleoside linkages contains a phosphorus atom. As used herein, oligonucleotides include oligonucleosides.

As used herein, "modified oligonucleotide" means an oligonucleotide comprising at least one modified nucleoside and/or at least one modified internucleoside linkage.

As used herein "internucleoside linkage" means a covalent linkage between adjacent nucleosides in an oligonucleotide.

As used herein "naturally occurring internucleoside linkage" means a 3' to 5' phosphodiester linkage.

As used herein, "modified internucleoside linkage" means any internucleoside linkage other than a naturally occurring internucleoside linkage.

As used herein, "oligomeric compound" means a polymeric structure comprising two or more substructures. In certain embodiments, an oligomeric compound comprises an oligonucleotide. In certain embodiments, an oligomeric compound comprises one or more conjugate groups and/or terminal groups. In certain embodiments, an oligomeric compound consists of an oligonucleotide.

As used herein, "terminal group" means one or more atom attached to either, or both, the 3 ' end or the 5' end of an oligonucleotide. In certain embodiments a terminal group is a conjugate group. In certain embodiments, a terminal group comprises one or more terminal group nucleosides.

As used herein, "conjugate" means an atom or group of atoms bound to an oligonucleotide or oligomeric compound. In general, conjugate groups modify one or more properties of the compound to which they are attached, including, but not limited to pharmacodynamic, pharmacokinetic, binding, absorption, cellular distribution, cellular uptake, charge and/or clearance properties.

As used herein, "conjugate linking group" means any atom or group of atoms used to attach a conjugate to an oligonucleotide or oligomeric compound.

As used herein, "antisense compound" means a compound comprising or consisting of an oligonucleotide at least a portion of which is complementary to a target nucleic acid to which it is capable of hybridizing, resulting in at least one antisense activity.

As used herein, "antisense activity" means any detectable and/or measurable change attributable to the hybridization of an antisense compound to its target nucleic acid.

As used herein, "detecting" or "measuring" means that a test or assay for detecting or measuring is performed. Such detection and/or measuring may result in a value of zero. Thus, if a test for detection or measuring results in a finding of no activity (activity of zero), the step of detecting or measuring the activity has nevertheless been performed.

As used herein, "detectable and/or measureable activity" means a measurable activity that is not zero.

As used herein, "essentially unchanged" means little or no change in a particular parameter, particularly relative to another parameter which changes much more. In certain embodiments, a parameter is essentially unchanged when it changes less than 5%. In certain embodiments, a parameter is essentially unchanged if it changes less than two-fold while another parameter changes at least ten- fold. For example, in certain embodiments, an antisense activity is a change in the amount of a target nucleic acid. In certain such embodiments, the amount of a non-target nucleic acid is essentially unchanged if it changes much less than the target nucleic acid does, but the change need not be zero.

As used herein, "expression" means the process by which a gene ultimately results in a protein. Expression includes, but is not limited to, transcription, post-transcriptional modification (e.g., splicing, polyadenlyation, addition of 5 '-cap), and translation.

As used herein, "target nucleic acid" means a nucleic acid molecule to which an antisense compound hybridizes.

As used herein, "single nucleotide polymorphism" or "SNP" means a single nucleotide variation between the genomes of individuals of the same species. In some cases, a SNP may be a single nucleotide deletion or insertion.

As used herein, "mRNA" means an RNA molecule that encodes a protein.

As used herein, "pre -mRNA" means an RNA transcript that has not been fully processed into mRNA. Pre-RNA includes one or more intron.

As used herein, "object RNA" means an RNA molecule other than a target RNA, the amount, activity, splicing, and/or function of which is modulated, either directly or indirectly, by a target nucleic acid. In certain embodiments, a target nucleic acid modulates splicing of an object RNA. In certain such embodiments, an antisense compound modulates the amount or activity of the target nucleic acid, resulting in a change in the splicing of an object RNA and ultimately resulting in a change in the activity or function of the object RNA.

As used herein, "microRNA" means a naturally occurring, small, non-coding RNA that represses gene expression of at least one mRNA. In certain embodiments, a microRNA represses gene expression by binding to a target site within a 3 ' untranslated region of an mRNA. In certain embodiments, a microRNA has a nucleobase sequence as set forth in miRBase, a database of published microRNA sequences found at http://microrna.sanger.ac.uk/sequences/. In certain embodiments, a microRNA has a nucleobase sequence as set forth in miRBase version 12.0 released September 2008, which is herein incorporated by reference in its entirety.

As used herein, "microRNA mimic" means an oligomeric compound having a sequence that is at least partially identical to that of a microRNA. In certain embodiments, a microRNA mimic comprises the microRNA seed region of a microRNA. In certain embodiments, a microRNA mimic modulates translation of more than one target nucleic acids. In certain embodiments, a microRNA mimic is double-stranded.

As used herein, "targeting" or "targeted to" means the association of an antisense compound to a particular target nucleic acid molecule or a particular region of a target nucleic acid molecule. An antisense compound targets a target nucleic acid if it is sufficiently complementary to the target nucleic acid to allow hybridization under physiological conditions.

As used herein, "nucleobase complementarity" or "complementarity" when in reference to nucleobases means a nucleobase that is capable of base pairing with another nucleobase. For example, in DNA, adenine (A) is complementary to thymine (T). For example, in RNA, adenine (A) is complementary to uracil (U). In certain embodiments, complementary nucleobase means a nucleobase of an antisense compound that is capable of base pairing with a nucleobase of its target nucleic acid. For example, if a nucleobase at a certain position of an antisense compound is capable of hydrogen bonding with a nucleobase at a certain position of a target nucleic acid, then the position of hydrogen bonding between the

oligonucleotide and the target nucleic acid is considered to be complementary at that nucleobase pair.

Nucleobases comprising certain modifications may maintain the ability to pair with a counterpart nucleobase and thus, are still capable of nucleobase complementarity.

As used herein, "non-complementary" in reference to nucleobases means a pair of nucleobases that do not form hydrogen bonds with one another.

As used herein, "complementary" in reference to oligomeric compounds (e.g., linked nucleosides, oligonucleotides, or nucleic acids) means the capacity of such oligomeric compounds or regions thereof to hybridize to another oligomeric compound or region thereof through nucleobase complementarity under stringent conditions. Complementary oligomeric compounds need not have nucleobase complementarity at each nucleoside. Rather, some mismatches are tolerated. In certain embodiments, complementary oligomeric compounds or regions are complementary at 70% of the nucleobases (70% complementary). In certain embodiments, complementary oligomeric compounds or regions are 80% complementary. In certain embodiments, complementary oligomeric compounds or regions are 90% complementary. In certain embodiments, complementary oligomeric compounds or regions are 95% complementary. In certain embodiments, complementary oligomeric compounds or regions are 100% complementary.

As used herein, "hybridization" means the pairing of complementary oligomeric compounds (e.g., an antisense compound and its target nucleic acid). While not limited to a particular mechanism, the most common mechanism of pairing involves hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases.

As used herein, "specifically hybridizes" means the ability of an oligomeric compound to hybridize to one nucleic acid site with greater affinity than it hybridizes to another nucleic acid site. In certain embodiments, an antisense oligonucleotide specifically hybridizes to more than one target site.

As used herein, "fully complementary" in reference to an oligonucleotide or portion thereof means that each nucleobase of the oligonucleotide or portion thereof is capable of pairing with a nucleobase of a complementary nucleic acid or contiguous portion thereof. Thus, a fully complementary region comprises no mismatches or unhybridized nucleobases in either strand.

As used herein, "percent complementarity" means the percentage of nucleobases of an oligomeric compound that are complementary to an equal-length portion of a target nucleic acid. Percent

complementarity is calculated by dividing the number of nucleobases of the oligomeric compound that are complementary to nucleobases at corresponding positions in the target nucleic acid by the total length of the oligomeric compound.

As used herein, "percent identity" means the number of nucleobases in a first nucleic acid that are the same type (independent of chemical modification) as nucleobases at corresponding positions in a second nucleic acid, divided by the total number of nucleobases in the first nucleic acid.

As used herein, "modulation" means a change of amount or quality of a molecule, function, or activity when compared to the amount or quality of a molecule, function, or activity prior to modulation. For example, modulation includes the change, either an increase (stimulation or induction) or a decrease

(inhibition or reduction) in gene expression. As a further example, modulation of expression can include a change in splice site selection of pre-mRNA processing, resulting in a change in the absolute or relative amount of a particular splice-variant compared to the amount in the absence of modulation.

As used herein, "motif means a pattern of chemical modifications in an oligomeric compound or a region thereof. Motifs may be defined by modifications at certain nucleosides and/or at certain linking groups of an oligomeric compound.

As used herein, "nucleoside motif means a pattern of nucleoside modifications in an oligomeric compound or a region thereof. The linkages of such an oligomeric compound may be modified or unmodified. Unless otherwise indicated, motifs herein describing only nucleosides are intended to be nucleoside motifs. Thus, in such instances, the linkages are not limited.

As used herein, "sugar motif means a pattern of sugar modifications in an oligomeric compound or a region thereof.

As used herein, "linkage motif means a pattern of linkage modifications in an oligomeric compound or region thereof. The nucleosides of such an oligomeric compound may be modified or unmodified. Unless otherwise indicated, motifs herein describing only linkages are intended to be linkage motifs. Thus, in such instances, the nucleosides are not limited.

As used herein, "nucleobase modification motif means a pattern of modifications to nucleobases along an oligonucleotide. Unless otherwise indicated, a nucleobase modification motif is independent of the nucleobase sequence.

As used herein, "sequence motif means a pattern of nucleobases arranged along an oligonucleotide or portion thereof. Unless otherwise indicated, a sequence motif is independent of chemical modifications and thus may have any combination of chemical modifications, including no chemical modifications.

As used herein, "type of modification" in reference to a nucleoside or a nucleoside of a "type" means the chemical modification of a nucleoside and includes modified and unmodified nucleosides. Accordingly, unless otherwise indicated, a "nucleoside having a modification of a first type" may be an unmodified nucleoside.

As used herein, "differently modified" mean chemical modifications or chemical substituents that are different from one another, including absence of modifications. Thus, for example, a MOE nucleoside and an unmodified DNA nucleoside are "differently modified," even though the DNA nucleoside is unmodified. Likewise, DNA and RNA are "differently modified," even though both are naturally-occurring unmodified nucleosides. Nucleosides that are the same but for comprising different nucleobases are not differently modified. For example, a nucleoside comprising a 2'-OMe modified sugar and an unmodified adenine nucleobase and a nucleoside comprising a 2'-OMe modified sugar and an unmodified thymine nucleobase are not differently modified.

As used herein, "the same type of modifications" refers to modifications that are the same as one another, including absence of modifications. Thus, for example, two unmodified DNA nucleoside have "the same type of modification," even though the DNA nucleoside is unmodified. Such nucleosides having the same type modification may comprise different nucleobases.

As used herein, "separate regions" means portions of an oligonucleotide wherein the chemical modifications or the motif of chemical modifications of any neighboring portions include at least one difference to allow the separate regions to be distinguished from one another.

As used herein, "pharmaceutically acceptable carrier or diluent" means any substance suitable for use in administering to an animal. In certain embodiments, a pharmaceutically acceptable carrier or diluent is sterile saline. In certain embodiments, such sterile saline is pharmaceutical grade saline.

As used herein, "substituent" and "substituent group," means an atom or group that replaces the atom or group of a named parent compound. For example a substituent of a modified nucleoside is any atom or group that differs from the atom or group found in a naturally occurring nucleoside (e.g., a modified 2'- substuent is any atom or group at the 2 '-position of a nucleoside other than H or OH). Substituent groups can be protected or unprotected. In certain embodiments, compounds of the present invention have substituents at one or at more than one position of the parent compound. Substituents may also be further substituted with other substituent groups and may be attached directly or via a linking group such as an alkyl or hydrocarbyl group to a parent compound.

Likewise, as used herein, "substituent" in reference to a chemical functional group means an atom or group of atoms differs from the atom or a group of atoms normally present in the named functional group. In certain embodiments, a substituent replaces a hydrogen atom of the functional group (e.g., in certain embodiments, the substituent of a substituted methyl group is an atom or group other than hydrogen which replaces one of the hydrogen atoms of an unsubstituted methyl group). Unless otherwise indicated, groups amenable for use as substituents include without limitation, halogen, hydroxyl, alkyl, alkenyl, alkynyl, acyl (- C(0)Raa), carboxyl (-C(0)0-Raa), aliphatic groups, alicyclic groups, alkoxy, substituted oxy (-O-Raa), aryl, aralkyl, heterocyclic radical, heteroaryl, heteroarylalkyl, amino (-N(Rbb)(RcC)), imino(=NRbb), amido

(-C(0)N(Rbb)(Rcc) or -N(Rbb)C(0)Raa), azido (-N3), nitro (-N02), cyano (-CN), carbamido

(-OC(0)N(Rbb)(Rcc) or -N(Rbb)C(0)ORaa), ureido (-N(Rbb)C(0)N(Rbb)(Rcc)), thioureido (-N(Rbb)C(S)N(Rbb)- (Rcc)), guanidinyl (-N(Rbb)C(=NRbb)N(Rbb)(Rcc)), amidinyl (-C(=NRbb)N(Rbb)(Rcc) or -N(Rbb)C(=NRbb)(Raa)), thiol (-SRbb), sulfmyl (-S(0)Rbb), sulfonyl (-S(0)2Rbb) and sulfonamidyl (-S(0)2N(Rbb)(Rcc) or -N(Rbb)S- (0)2Rbb). Wherein each Raa, Rbb and Rcc is, independently, H, an optionally linked chemical functional group or a further substituent group with a preferred list including without limitation, alkyl, alkenyl, alkynyl, aliphatic, alkoxy, acyl, aryl, aralkyl, heteroaryl, alicyclic, heterocyclic and heteroarylalkyl. Selected substituents within the compounds described herein are present to a recursive degree.

As used herein, "alkyl," as used herein, means a saturated straight or branched hydrocarbon radical containing up to twenty four carbon atoms. Examples of alkyl groups include without limitation, methyl, ethyl, propyl, butyl, isopropyl, n-hexyl, octyl, decyl, dodecyl and the like. Alkyl groups typically include from 1 to about 24 carbon atoms, more typically from 1 to about 12 carbon atoms (C1-C12 alkyl) with from 1 to about 6 carbon atoms being more preferred.

As used herein, "alkenyl," means a straight or branched hydrocarbon chain radical containing up to twenty four carbon atoms and having at least one carbon-carbon double bond. Examples of alkenyl groups include without limitation, ethenyl, propenyl, butenyl, 1 -methyl-2-buten- 1 -yl, dienes such as 1,3-butadiene and the like. Alkenyl groups typically include from 2 to about 24 carbon atoms, more typically from 2 to about 12 carbon atoms with from 2 to about 6 carbon atoms being more preferred. Alkenyl groups as used herein may optionally include one or more further substituent groups.

As used herein, "alkynyl," means a straight or branched hydrocarbon radical containing up to twenty four carbon atoms and having at least one carbon-carbon triple bond. Examples of alkynyl groups include, without limitation, ethynyl, 1-propynyl, 1-butynyl, and the like. Alkynyl groups typically include from 2 to about 24 carbon atoms, more typically from 2 to about 12 carbon atoms with from 2 to about 6 carbon atoms being more preferred. Alkynyl groups as used herein may optionally include one or more further substituent groups.

As used herein, "acyl," means a radical formed by removal of a hydroxyl group from an organic acid and has the general Formula -C(0)-X where X is typically aliphatic, alicyclic or aromatic. Examples include aliphatic carbonyls, aromatic carbonyls, aliphatic sulfonyls, aromatic sulfinyls, aliphatic sulfinyls, aromatic phosphates, aliphatic phosphates and the like. Acyl groups as used herein may optionally include further substituent groups.

As used herein, "alicyclic" means a cyclic ring system wherein the ring is aliphatic. The ring system can comprise one or more rings wherein at least one ring is aliphatic. Preferred alicyclics include rings having from about 5 to about 9 carbon atoms in the ring. Alicyclic as used herein may optionally include further substituent groups. As used herein, "aliphatic" means a straight or branched hydrocarbon radical containing up to twenty four carbon atoms wherein the saturation between any two carbon atoms is a single, double or triple bond. An aliphatic group preferably contains from 1 to about 24 carbon atoms, more typically from 1 to about 12 carbon atoms with from 1 to about 6 carbon atoms being more preferred. The straight or branched chain of an aliphatic group may be interrupted with one or more heteroatoms that include nitrogen, oxygen, sulfur and phosphorus. Such aliphatic groups interrupted by heteroatoms include without limitation, polyalkoxys, such as polyalkylene glycols, polyamines, and polyimines. Aliphatic groups as used herein may optionally include further substituent groups.

As used herein, "alkoxy" means a radical formed between an alkyl group and an oxygen atom wherein the oxygen atom is used to attach the alkoxy group to a parent molecule. Examples of alkoxy groups include without limitation, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n- pentoxy, neopentoxy, n-hexoxy and the like. Alkoxy groups as used herein may optionally include further substituent groups.

As used herein, "aminoalkyl" means an amino substituted C1-C12 alkyl radical. The alkyl portion of the radical forms a covalent bond with a parent molecule. The amino group can be located at any position and the aminoalkyl group can be substituted with a further substituent group at the alkyl and/or amino portions.

As used herein, "aralkyl" and "arylalkyl" mean an aromatic group that is covalently linked to a C1-C12 alkyl radical. The alkyl radical portion of the resulting aralkyl (or arylalkyl) group forms a covalent bond with a parent molecule. Examples include without limitation, benzyl, phenethyl and the like. Aralkyl groups as used herein may optionally include further substituent groups attached to the alkyl, the aryl or both groups that form the radical group.

As used herein, "aryl" and "aromatic" mean a mono- or polycyclic carbocyclic ring system radicals having one or more aromatic rings. Examples of aryl groups include without limitation, phenyl, naphthyl, tetrahydronaphthyl, indanyl, idenyl and the like. Preferred aryl ring systems have from about 5 to about 20 carbon atoms in one or more rings. Aryl groups as used herein may optionally include further substituent groups.

As used herein, "halo" and "halogen," mean an atom selected from fluorine, chlorine, bromine and iodine.

As used herein, "heteroaryl," and "heteroaromatic," mean a radical comprising a mono- or polycyclic aromatic ring, ring system or fused ring system wherein at least one of the rings is aromatic and includes one or more heteroatoms. Heteroaryl is also meant to include fused ring systems including systems where one or more of the fused rings contain no heteroatoms. Heteroaryl groups typically include one ring atom selected from sulfur, nitrogen or oxygen. Examples of heteroaryl groups include without limitation, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl and the like. Heteroaryl radicals can be attached to a parent molecule directly or through a linking moiety such as an aliphatic group or hetero atom. Heteroaryl groups as used herein may optionally include further substituent groups.

Oligomeric Compounds

In certain embodiments, the present invention provides oligomeric compounds. In certain embodiments, such oligomeric compounds comprise oligonucleotides optionally comprising one or more conjugate and/or terminal groups. In certain embodiments, an oligomeric compound consists of an oligonucleotide. In certain embodiments, oligonucleotides comprise one or more chemical modifications. Such chemical modifications include modifications one or more nucleoside (including modifications to the sugar moiety and/or the nucleobase) and/or modifications to one or more internucleoside linkage.

Certain Sugar Moieties

In certain embodiments, oligomeric compounds of the invention comprise one or more modifed nucleosides comprising a modifed sugar moiety. Such oligomeric compounds comprising one or more sugar- modified nucleosides may have desirable properties, such as enhanced nuclease stability or increased binding affinity with a target nucleic acid relative to oligomeric compounds comprising only nucleosides comprising naturally occurring sugar moieties. In certain embodiments, modified sugar moieties are substitued sugar moieties. In certain embodiments, modified sugar moieties are sugar surrogates. Such sugar surogates may comprise one or more substitutions corresponding to those of substituted sugar moieties.

In certain embodiments, modified sugar moieties are substituted sugar moieties comprising one or more non-bridging sugar substituent, including but not limited to substituents at the 2' and/or 5' positions. Examples of sugar substituents suitable for the 2 '-position, include, but are not limited to: 2'-F, 2'-OCH3 ("OMe" or "O-methyl"), and 2'-0(CH2)2OCH3 ("MOE"). In certain embodiments, sugar substituents at the 2' position is selected from allyl, amino, azido, thio, O-allyl, O-Ci-Cio alkyl, O-CpCio substituted alkyl; OCF3, 0(CH2)2SCH3, 0(CH2)2-0-N(Rm)(Rn), and 0-CH2-C(=0)-N(Rm)(Rn), where each Rm and Rn is, independently, H or substituted or unsubstituted Ci-Cio alkyl. Examples of sugar substituents at the 5'- position, include, but are not limited to:, 5'-methyl (R or S); 5'-vinyl, and 5'-methoxy. In certain

embodiments, substituted sugars comprise more than one non-bridging sugar substituent, for example, 2'-F- 5'-methyl sugar moieties (see,c.g., PCT International Application WO 2008/101 157, for additional 5', 2'-bis substituted sugar moieties and nucleosides).

Nucleosides comprising 2' -substituted sugar moieties are referred to as 2 '-substituted nucleosides. In certain embodiments, a 2'- substituted nucleoside comprises a 2'-substituent group selected from halo, allyl, amino, azido, SH, CN, OCN, CF3, OCF3, O, S, or N(Rm)-alkyl; O, S, or N(Rm)-alkenyl; O, S or N(Rm)- alkynyl; O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, 0(CH2)2SCH3, 0-(CH2)2-0- N(Rm)(Rn) or 0-CH2-C(=0)-N(Rm)(Rn), where each Rm and Rn is, independently, H, an amino protecting group or substituted or unsubstituted Ci-Cio alkyl. These 2'-substituent groups can be further substituted with one or more substituent groups independently selected from hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO2), thiol, thioalkoxy (S-alkyl), halogen, alkyl, aryl, alkenyl and alkynyl.

In certain embodiments, a 2'- substituted nucleoside comprises a 2 '-substituent group selected from F, NH2, N3, OCF3; O-CH3, 0(CH2)3NH2, CH2-CH=CH2, 0-CH2-CH=CH2, OCH2CH2OCH3, 0(CH2)2SCH3, 0-(CH2)2-0-N(Rm)(Rn), 0(CH2)20(CH2)2N(CH3)2, and N-substituted acetamide (0-CH2-C(=0)-N(Rm)(Rn) where each Rm and Rn is, independently, H, an amino protecting group or substituted or unsubstituted CpCio alkyl.

In certain embodiments, a 2'- substituted nucleoside comprises a sugar moiety comprising a 2'- substituent group selected from F, OCF3, 0-CH3, OCH2CH2OCH3, 0(CH2)2SCH3, 0-(CH2)2-0-N(CH3)2, -0(CH2)20(CH2)2N(CH3)2, and 0-CH2-C(=0)-N(H)CH3.

In certain embodiments, a 2'- substituted nucleoside comprises a sugar moiety comprising a 2'- substituent group selected from F, 0-CH3, and OCH2CH2OCH3.

Certain modifed sugar moieties comprise a bridging sugar substituent that forms a second ring resulting in a bicyclic sugar moiety. In certain such embodiments, the bicyclic sugar moiety comprises a bridge between the 4' and the 2' furanose ring atoms. Examples of such 4' to 2' sugar substituents, include, but are not limited to: -[C(Ra)(Rb)]n-, -[C(Ra)(Rb)]n-0-, -C(RaRb)-N(R)-0- or, -C(RaRb)-0-N(R)-; 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2',. 4'-(CH2)-0-2' (LNA); 4'-(CH2)-S-2'; 4'-(CH2)2-0-2' (ENA); 4'-CH(CH3)-0-2' (cEt) and 4'-CH(CH2OCH3)-0-2',and analogs thereof (see, e.g., U.S. Patent 7,399,845, issued on July 15, 2008); 4'-C(CH3)(CH3)-0-2'and analogs thereof, (see, e.g., WO2009/006478, published January 8, 2009); 4'- CH2-N(OCH3)-2' and analogs thereof (see, e.g., WO2008/150729, published December 1 1, 2008); 4'-CH2-0- N(CH3)-2' (see, e.g., US2004/0171570, published September 2, 2004 ); 4'-CH2-0-N(R)-2', and 4'-CH2-N(R)- 0-2'-, wherein each Ris, independently, H, a protecting group, or CrCi2 alkyl; 4'-CH2-N(R)-0-2', wherein R is H, C1-C12 alkyl, or a protecting group (see, U.S. Patent 7,427,672, issued on September 23, 2008); 4'-CH2- C(H)(CH3)-2' (see, e.g., Chattopadhyaya, et al, J. Org. Chem.,2009, 74, 1 18- 134); and 4'-CH2-C(=CH2)-2' and analogs thereof (see, published PCT International Application WO 2008/154401, published on December 8, 2008).

In certain embodiments, such 4' to 2' bridges independently comprise from 1 to 4 linked groups independently selected from -[C(Ra)(Rb)]n-, -C(Ra)=C(Rb)-, -C(Ra)=N-, -C(=NRa)-, -C(=0)-, -C(=S)-, -0-, - Si(Ra)2-, -S(=0)x-, and -N(Ra)-;

wherein:

x is 0, 1, or 2;

n is 1, 2, 3, or 4;

each Ra and Rb is, independently, H, a protecting group, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, C2-Ci2 alkenyl, substituted C2-Ci2 alkenyl, C2-Ci2 alkynyl, substituted C2-Ci2 alkynyl, C5-C2o aryl, substituted C5-C2o aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJi, NJiJ2, SJi, N3, COOJi, acyl (C(=0)- H), substituted acyl, CN, sulfonyl (S(=0)2-Ji), or sulfoxyl and

each Ji and J2 is, independently, H, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, acyl (C(=0)- H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C1-Q2 aminoalkyl, substituted C1-C12 aminoalkyl, or a protecting group.

Nucleosides comprising bicyclic sugar moieties are referred to as bicyclic nucleosides or BNAs. Bicyclic nucleosides include, but are not limited to, (A) a-L-Methyleneoxy (4'-CH2-0-2') BNA , (B) β-D- Methyleneoxy (4'-CH2-0-2') BNA (also referred to as locked nucleic acid or LNA) , (C) Ethyleneoxy (4'- (CH2)2-0-2') BNA , (D) Aminooxy (4'-CH2-0-N(R)-2') BNA, (E) Oxyamino (4'-CH2-N(R)-0-2') BNA, (F) Methyl(methyleneoxy) (4'-CH(CH3)-0-2') BNA (also referred to as constrained ethyl or cEt), (G) methylene-thio (4'-CH2-S-2') BNA, (H) methylene-amino (4'-CH2-N(R)-2') BNA, (I) methyl carbocyclic (4'-CH2-CH(CH3)-2') BNA, and (J) propylene carbocyclic (4'-(CH2)3-2') BNA as depicted below.

Figure imgf000045_0001

(A) (B) (C)

Figure imgf000045_0002
wherein Bx is a nucleobase moiety and R is, independently, H, a protecting group, or C1-C12 alkyl.

Additional bicyclic sugar moieties are known in the art, for example: Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Wahlestedt et al., Proc. Natl. Acad. Sci. U. S. A. , 2000, 97, 5633-5638; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035- 10039; Srivastava et al., J. Am. Chem. Soc, 129(26) 8362-8379 (Jul. 4, 2007); Elayadi et al, Curr. Opinion Invens. Drugs, 2001 , 2, 558-561 ; Braasch et al., Chem. Biol., 2001 , 8, 1 -7; Orum et al., Curr. Opinion Mol. Ther., 2001 , 3, 239-243; U.S. Patent Nos. 7,053,207, 6,268,490, 6,770,748, 6,794,499, 7,034, 133, 6,525, 191 , 6,670,461 , and 7,399,845; WO 2004/106356, WO 1994/14226, WO 2005/021570, and WO 2007/134181 ; U.S. Patent Publication Nos. US2004/0171570, US2007/0287831 , and US2008/0039618; U.S. Patent Serial Nos. 12/129, 154, 60/989,574, 61/026,995, 61/026,998, 61/056,564, 61/086,231 , 61/097,787, and 61/099,844; and PCT International Applications Nos. PCT/US2008/064591 , PCT/US2008/066154, and PCT/US2008/068922.

In certain embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by isomeric configuration. For example, a nucleoside comprising a 4' -2' methylene-oxy bridge, may be in the a-L configuration or in the β-D configuration. Previously, a-L- methyleneoxy (4'-CH2-0-2') bicyclic nucleosides have been incorporated into antisense oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372).

In certain embodiments, substituted sugar moieties comprise one or more non-bridging sugar substituent and one or more bridging sugar substituent (e.g., 5 '-substituted and 4'-2' bridged sugars), {see, PCT International Application WO 2007/134181 , published on 1 1/22/07, wherein LNA is substituted with, for example, a 5'-methyl or a 5'-vinyl group).

In certain embodiments, modified sugar moieties are sugar surrogates. In certain such embodiments, the oxygen atom of the naturally occuring sugar is substituted, e.g., with a sulfer, carbon or nitrogen atom. In certain such embodiments, such modified sugar moiety also comprises bridging and/or non-bridging substituents as described above. For example, certain sugar surogates comprise a 4 '-sulfer atom and a substitution at the 2'-position (see,c.g., published U.S. Patent Application US2005/0130923, published on June 16, 2005) and/or the 5' position. By way of additional example, carbocyclic bicyclic nucleosides having a 4'-2' bridge have been described (see, e.g., Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443 and Albaek ei a/., J. Org. Chem., 2006, 71, 7731 -7740).

In certain embodiments, sugar surrogates comprise rings having other than 5-atoms. For example, in certain embodiments, a sugar surrogate comprises a six-membered tetrahydropyran. Such tetrahydropyrans may be further modified or substituted. Nucleosides comprising such modified tetrahydropyrans include, but are not limited to, hexitol nucleic acid (UNA), anitol nucleic acid (ANA), manitol nucleic acid (MNA) (see Leumann, CJ. Bioorg. & Med. Chem. (2002) 10:841 -854), fluoro UNA (F-HNA), and those compounds having Formula VII:

Figure imgf000047_0001

VII

wherein independently for each of said at least one tetrahydropyran nucleoside analog of Formula VII:

Bx is a nucleobase moiety;

T3 and T are each, independently, an internucleoside linking group linking the tetrahydropyran nucleoside analog to the antisense compound or one of T3 and T4 is an internucleoside linking group linking the tetrahydropyran nucleoside analog to the antisense compound and the other of T3 and T is H, a hydroxyl protecting group, a linked conjugate group, or a 5' or 3'-terminal group;

qi, q2, q3, q4, qs, q6 ¾nd q7 are each, independently, H, Ci-Ce alkyl, substituted Ci-Ce alkyl, C2-C6 alkenyl, substituted C2-C6 alkenyl, C2-C6 alkynyl, or substituted C2-C6 alkynyl; and

one of Ri and R2 is hydrogen and the other is selected from halogen, substituted or unsubstituted alkoxy, NJiJ2, SJb N3, OC(=X)Jb OC(=X)NJ!J2, NJ3C(=X)NJ!J2, and CN, wherein X is O, S or NJb and each Ji, J 2, and J3 is, independently, H or Ci-C6 alkyl.

In certain embodiments, the modified THP nucleosides of Formula VII are provided wherein qb q2, q3, q4, q5, q6 and q7 are each H. In certain embodiments, at least one of q q2, q3, q4, qs, q6 and q7 is other than H. In certain embodiments, at least one of qb q2, q3, q4, qs, q6 and q7 is methyl. In certain embodiments, THP nucleosides of Formula VII are provided wherein one of Ri and R2 is F. In certain embodiments, Ri is fluoro and R2 is H, Ri is methoxy and R2 is H, and Ri is methoxyethoxy and R2 is H.

Many other bicyclo and tricyclo sugar surrogate ring systems are also known in the art that can be used to modify nucleosides for incorporation into antisense compounds (see, e.g., review article: Leumann, J. C, Bioorganic & Medicinal Chemistry, 2002, 10, 841 -854).

Combinations of modifications are also provided without limitation, such as 2'-F-5'-methyl substituted nucleosides (see PCT International Application WO 2008/101 157 Published on 8/21/08 for other disclosed 5', 2'-bis substituted nucleosides) and replacement of the ribosyl ring oxygen atom with S and further substitution at the 2'-position (see published U.S. Patent Application US2005-0130923, published on June 16, 2005) or alternatively 5'-substitution of a bicyclic nucleic acid (see PCT International Application WO 2007/134181 , published on 1 1/22/07 wherein a 4'-CH2-0-2' bicyclic nucleoside is further substituted at the 5' position with a 5'-methyl or a 5'-vinyl group). The synthesis and preparation of carbocyclic bicyclic nucleosides along with their oligomerization and biochemical studies have also been described (see, e.g., Srivastava et al, J. Am. Chem. Soc. 2007, 129(26), 8362-8379). Certain Nucleobases

In certain embodiments, nucleosides of the present invention comprise one or more unmodified nucleobases. In certain embodiments, nucleosides of the present invention comprise one or more modifed nucleobases.

In certain embodiments, modified nucleobases are selected from: universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases as defined herein. 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2-aminopropyladenine, 5- propynyluracil; 5-propynylcytosine; 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6- methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl (-C≡C- CH3) uracil and cytosine and other alkynyl derivatives of pyrimidine bases, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8- substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 2-F-adenine, 2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine, 3-deazaguanine and 3-deazaadenine, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases as defined herein. Further modified nucleobases include tricyclic pyrimidines such as phenoxazine cytidine( [5,4-b][l,4]benzoxazin- 2(3H)-one), phenothiazine cytidine (lH-pyrimido[5,4-b] [l,4]benzothiazin-2(3H)-one), G-clamps such as a substituted phenoxazine cytidine (e.g. 9-(2-aminoethoxy)-H-pyrimido[5,4-b][l,4]benzoxazin-2(3H)-one), carbazole cytidine (2H-pyrimido[4,5-b]indol-2-one), pyridoindole cytidine (H-pyrido[3',2':4,5]pyrrolo[2,3- d]pyrimidin-2-one). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2- pyridone. Further nucleobases include those disclosed in United States Patent No. 3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, J.I., Ed., John Wiley & Sons, 1990, 858-859; those disclosed by Englisch et al, Angewandte Chemie, International Edition, 1991, 30, 613; and those disclosed by Sanghvi, Y.S., Chapter 15, Antis ens e Research and Applications , Crooke, S.T. and Lebleu, B., Eds., CRC Press, 1993, 273-288.

Representative United States patents that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include without limitation, U.S. 3,687,808; 4,845,205; 5, 130,302; 5,134,066; 5, 175,273; 5,367,066; 5,432,272; 5,457, 187; 5,459,255; 5,484,908; 5,502,177;

5,525,71 1 ; 5,552,540; 5,587,469; 5,594, 121 ; 5,596,091 ; 5,614,617; 5,645,985; 5,681,941 ; 5,750,692;

5,763,588; 5,830,653 and 6,005,096, certain of which are commonly owned with the instant application, and each of which is herein incorporated by reference in its entirety.

Certain Internucleoside Linkages In certain embodiments, the present invention provides oligomeric compounds comprising linked nucleosides. In such embodiments, nucleosides may be linked together using any internucleoside linkage. The two main classes of internucleoside linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus containing internucleoside linkages include, but are not limited to, phosphodiesters (P=0), phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates (P=S). Representative non-phosphorus containing internucleoside linking groups include, but are not limited to, methylenemethylimino (-CH2-N(CH3)-0-CH2-), thiodiester (-O-C(O)-S-), thionocarbamate (-0- C(0)(NH)-S-); siloxane (-0-Si(H)2-0-); and Ν,Ν'-dimethylhydrazine (-CH2-N(CH3)-N(CH3)-). Modified linkages, compared to natural phosphodiester linkages, can be used to alter, typically increase, nuclease resistance of the oligomeric compound. In certain embodiments, internucleoside linkages having a chiral atom can be prepared as a racemic mixture, or as separate enantiomers. Representative chiral linkages include, but are not limited to, alkylphosphonates and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorous-containing internucleoside linkages are well known to those skilled in the art.

The oligonucleotides described herein contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric configurations that may be defined, in terms of absolute stereochemistry, as (R) or (S), a or β such as for sugar anomers, or as (D) or (L) such as for amino acids etc. Included in the antisense compounds provided herein are all such possible isomers, as well as their racemic and optically pure forms.

Neutral internucleoside linkages include without limitation, phosphotriesters, methylphosphonates, MMI (3'-CH2-N(CH3)-0-5'), amide-3 (3'-CH2-C(=0)-N(H)-5'), amide-4 (3'-CH2-N(H)-C(=0)-5'), formacetal (3'-0-CH2-0-5'), and thioformacetal (3'-S-CH2-0-5'). Further neutral internucleoside linkages include nonionic linkages comprising siloxane (dialkylsiloxane), carboxylate ester, carboxamide, sulfide, sulfonate ester and amides (See for example: Carbohydrate Modifications in Antisense Research; Y.S. Sanghvi and P.D. Cook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutral internucleoside linkages include nonionic linkages comprising mixed N, O, S and CH2 component parts.

Certain Motifs

In certain embodiments, the present invention provides oligomeric compounds comprising oligonucleotides. In certain embodiments, such oligonucleotides comprise one or more chemical modification. In certain embodiments, chemically modified oligonucleotides comprise one or more modified sugars. In certain embodiments, chemically modified oligonucleotides comprise one or more modified nucleobases. In certain embodiments, chemically modified oligonucleotides comprise one or more modified internucleoside linkages. In certain embodiments, the chemically modifications (sugar modifications, nucleobase modifications, and/or linkage modifications) define a pattern or motif. In certain embodiments, the patterns of chemical modifications of sugar moieties, internucleoside linkages, and nucleobases are each independent of one another. Thus, an oligonucleotide may be described by its sugar modification motif, internucleoside linkage motif and/or nucleobase modification motif (as used herein, nucleobase modification motif describes the chemical modifications to the nucleobases independent of the sequence of nucleobases).

Certain sugar motifs

In certain embodiments, oligonucleotides comprise one or more type of modified sugar moieties and/or naturally occurring sugar moieties arranged along an oligonucleotide or region thereof in a defined pattern or sugar modification motif. Such motifs may include any of the sugar modifications discussed herein and/or other known sugar modifications.

In certain embodiments, the oligonucleotides comprise or consist of a region having a gapmer sugar modification motif, which comprises two external regions or "wings" and an internal region or "gap." The three regions of a gapmer motif (the 5 '-wing, the gap, and the 3 '-wing) form a contiguous sequence of nucleosides wherein at least some of the sugar moieties of the nucleosides of each of the wings differ from at least some of the sugar moieties of the nucleosides of the gap. Specifically, at least the sugar moieties of the nucleosides of each wing that are closest to the gap (the 3 '-most nucleoside of the 5 '-wing and the 5 '-most nucleoside of the 3 '-wing) differ from the sugar moiety of the neighboring gap nucleosides, thus defining the boundary between the wings and the gap. In certain embodiments, the sugar moieties within the gap are the same as one another. In certain embodiments, the gap includes one or more nucleoside having a sugar moiety that differs from the sugar moiety of one or more other nucleosides of the gap. In certain embodiments, the sugar modification motifs of the two wings are the same as one another (symmetric gapmer). In certain embodiments, the sugar modification motifs of the 5'-wing differs from the sugar modification motif of the 3'-wing (asymmetric gapmer).

Certain 5 '-wings

In certain embodiments, the 5'- wing of a gapmer consists of 1 to 5 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 2 to 5 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 3 to 5 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 4 or 5 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 1 to 4 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 1 to 3 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 1 or 2 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 2 to 4 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 2 or 3 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 3 or 4 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 1 nucleoside. In certain embodiments, the 5'- wing of a gapmer consists of 2 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 31inked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 4 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer consists of 5 linked nucleosides. In certain embodiments, the 5'- wing of a gapmer comprises at least one bicyclic nucleoside. In certain embodiments, the 5'- wing of a gapmer comprises at least two bicyclic nucleosides. In certain embodiments, the 5'- wing of a gapmer comprises at least three bicyclic nucleosides. In certain

embodiments, the 5'- wing of a gapmer comprises at least four bicyclic nucleosides. In certain embodiments, the 5'- wing of a gapmer comprises at least one constrained ethyl nucleoside. In certain embodiments, the 5'- wing of a gapmer comprises at least one LNA nucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a bicyclic nucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a constrained ethyl nucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a LNA nucleoside.

In certain embodiments, the 5'- wing of a gapmer comprises at least one non-bicyclic modified nucleoside. In certain embodiments, the 5'- wing of a gapmer comprises at least one 2 '-substituted nucleoside. In certain embodiments, the 5'- wing of a gapmer comprises at least one 2'-MOE nucleoside. In certain embodiments, the 5'- wing of a gapmer comprises at least one 2'-OMe nucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a non-bicyclic modified nucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a 2 '-substituted nucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a 2'-MOE nucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a 2'-OMe nucleoside.

In certain embodiments, the 5'- wing of a gapmer comprises at least one 2'-deoxynucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a 2'-deoxynucleoside. In a certain embodiments, the 5'- wing of a gapmer comprises at least one ribonucleoside. In certain embodiments, each nucleoside of the 5'- wing of a gapmer is a ribonucleoside.

In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one non-bicyclic modified nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2 '-substituted nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2'-MOE nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2'-OMe nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2'-deoxynucleoside.

In certain embodiments, the 5 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one non-bicyclic modified nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2 '-substituted nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2'-MOE nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2'-OMe nucleoside. In certain embodiments, the 5'-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2'-deoxynucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one LNA nucleoside and at least one non-bicyclic modified nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'-substituted nucleoside. In certain embodiments, the 5'-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'-MOE nucleoside. In certain embodiments, the 5'-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'-OMe nucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'- deoxynucleoside.

In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one non-bicyclic modified nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5'- wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one non-bicyclic modified nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one LNA nucleoside, at least one non-bicyclic modified nucleoside, and at least one 2'- deoxynucleoside.

In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one 2 '-substituted nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one 2'-substituted nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one LNA nucleoside, at least one 2 '-substituted nucleoside, and at least one 2'-deoxynucleoside.

In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one 2'-MOE nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one 2'-MOE nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one LNA nucleoside, at least one 2'-MOE nucleoside, and at least one 2'-deoxynucleoside.

In certain embodiments, the 5 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one 2'-OMe nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5'-wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one 2'-OMe nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 5 '-wing of a gapmer comprises at least one LNA nucleoside, at least one 2'-OMe nucleoside, and at least one 2'-deoxynucleoside.

In certain embodiments, the 5'- wing of a gapmer has a sugar motif selected from among those listed in the following non-limiting table:

Table 1

Certain 5 '-Wing Sugar Motifs

Figure imgf000052_0001
a A-A-B 2d A-A-L 2g A-A-K

3a A-D-B 3d A-D-L 3g A-D-K a B-D-A 4d L-D-A 4g K-D-A

5a B-A-A 5d L-A-A 5g K-A-A

6a B-B-B 6d L-L-L 6g K-K-K

7a A-A-A 7d A-A-A 7g A-A-A

8a A-D-D-B 8d A-D-D-L 8g A-D-D-K

9a B-D-D-A 9d L-D-D-A 9g K-D-D-A

10a A-A-A-B lOd A-A-A-L 10g A-A-A-K

1 1a B-A-A-A l id L-A-A-A l lg K-A-A-A

12a A-A-A-A 12d A-A-A-A 12g A-A-A-A

13a B-D-D-B 13d L-D-D-L 13g K-D-D-K

14a A-A-A-A 14d A-A-A-A 14g A-A-A-A

15a B-B-B-B 15d L-L-L-L 15g K-K-K-K

16a A-A-A-A-A 16d A-A-A-A-A 16g A-A-A-A-A

17a A-D-A-D-B 17d A-D-A-D-L 17g A-D-A-D-K

18a A-D-B-D-A 18d A-D-L-D-A 18g A-D-K-D-A

19a B-D-A-D-A 19d L-D-A-D-A 19g K-D-A-D-A 0a A-A-A-A-B 20d A-A-A-A-L 20g A-A-A-A-K 1a A-A-B-A-A 21d A-A-L-A-A 21g A-A-K-A-A 2a B-A-A-A-A 22d L-A-A-A-A 22g K-A-A-A-A lb E-B-B le E-L-L lh E-K-K b E-E-B 2e E-E-L 2h E-E-K

3b E-D-B 3e E-D-L 3h E-D-K b B-D-E 4e L-D-E 4h K-D-E

5b B-E-E 5e L-E-E 5h K-E-E

6b B-B-B 6e L-L-L 6h K-K-K

7b E-E-E 7e E-E-E 7h E-E-E

8b E-D-D-B 8e E-D-D-L 8h E-D-D-K

9b B-D-D-E 9e L-D-D-E 9h K-D-D-E

10b E-E-E-B lOe E-E-E-L lOh E-E-E-K l ib B-E-E-E l ie L-E-E-E 1 lh K-E-E-E

12b E-E-E-E 12e E-E-E-E 12h E-E-E-E

13b B-D-D-B 13e L-D-D-L 13h K-D-D-K

14b E-E-E-E 14e E-E-E-E 14h E-E-E-E

15b B-B-B-B 15e L-L-L-L 15h K-K-K-K

16b E-E-E-E-E 16e E-E-E-E-E 16h E-E-E-E-E

17b E-D-E-D-B 17e E-D-E-D-L 17h E-D-E-D-K

18b E-D-B-D-E 18e E-D-L-D-E 18h E-D-K-D-E

19b B-D-E-D-E 19e L-D-E-D-E 19h K-D-E-D-E 0b E-E-E-E-B 20e E-E-E-E-L 20h E-E-E-E-K 1b E-E-B-E-E 21e E-E-L-E-E 21h E-E-K-E-E 2b B-E-E-E-E 22e L-E-E-E-E 22h K-E-E-E-E lc M-B-B If M-L-L li M-K-K 2c M-M-B 2f M-M-L 2i M-M-K

3c M-D-B 3f M-D-L 3i M-D-K

4c B-D-M 4f L-D-M 4i K-D-M

5c B-M-M 5f L-M-M 5i K-M-M

6c B-B-B 6f L-L-L 6i K-K-K

7c M-M-M 7f M-M-M 7i M-M-M

8c M-D-D-B 8f M-D-D-L 8i M-D-D-K

9c B-D-D-M 9f L-D-D-M 9i K-D-D-M

10c M-M-M-B lOf M-M-M-L lOi M-M-M-K

1 1c B-M-M-M 1 If L-M-M-M Hi K-M-M-M

12c M-M-M-M 12f M-M-M-M 12i M-M-M-M

13c B-D-D-B 13f L-D-D-L 13i K-D-D-K

14c M-M-M-M 14f M-M-M-M 14i M-M-M-M

15c B-B-B-B 15f L-L-L-L 15i K-K-K-K

16c M-M-M-M-M 16f M-M-M-M-M 16i M-M-M-M-M

17c M-D-M-D-B 17f M-D-M-D-L 17i M-D-M-D-K

18c M-D-B-D-M 18f M-D-L-D-M 18i M-D-K-D-M

19c B-D-M-D-M 19f L-D-M-D-M 19i K-D-M-D-M

20c M-M-M-M-B 20f M-M-M-M-L 20i M-M-M-M-K

21c M-M-B-M-M 21f M-M-L-M-M 21i M-M-K-M-M

22c B-M-M-M-M 22f L-M-M-M-M 22i K-M-M-M-M ij A-L-K lk A-K-L 11 E-L-K

¾ M-E-K 2k M-E-L 21 E-M-K

3j L-D-K 3k K-D-L 31 B-D-K

4j K-D-A 4k L-D-K 41 K-B-L

5j B-M-E 5k L-M-E 51 K-M-E

6j K-L-L 6k L-K-L 61 L-K-K

7j E-M-E 7k M-E-M 71 M-E-E

8j E-D-D-M 8k K-D-D-L 81 L-D-D-K

¾ M-D-D-E 9k L-D-K-E 91 K-D-L-E

10j E-M-E-B 10k E-M-E-L 101 E-M-E-K i ij B-E-E-M I lk L-E-E-M 1 11 K-E-E-M

1¾ E-E-E-M 12k M-E-E-E 121 E-M-E-E

1¾ K-L-D-K 13k L-K-D-L 131 K-D-L-K

14j E-M-E-M 14k M-EM-E 141 E-E-M-E

15j K-L-L-K 15k L-K-L-K 151 K-L-K-K

16j E-E-M-E-E 16k M-E-E-E-M 161 E-E-M-M-E

173 E-D-M-D-K 17k E-D-M-D-L 171 M-D-E-D-K

1 ¾ E-D-K-D-M 18k E-D-L-D-M 181 M-D-K-D-E

1¾ B-D-A-D-A 19k L-D-A-D-A 191 K-D-A-D-A

20j E-M-E-E-L 20k E-M-M-E-L 201 M-E-E-E-K

21j E-E-K-M-M 21k E-E-L-M-M 211 E-M-K-E-E

22j B-E-M-E-A 22k L-E-A-M-A 221 K-E-A-A-A

23j K-D-K-D-K 23k E-K-E-K-D-K In the above table, "A" represents a nucleoside comprising a 2'-substituted sugar moiety; "B" represents a bicyclic nucleoside; "D" represents a 2'-deoxynucleoside; "K" represents a constrained ethyl nucleoside; "L" represents an LNA nucleoside; "E" represents a 2'-MOE nucleoside; and "M" represents a 2 '-OMe nucleoside.

In certain embodiments, an oligonucleotide comprises any 5 '-wing motif provided herein. In certain such embodiments, the oligonucleotide is a 5'-hemimer (does not comprise a 3 '-wing). In certain embodiments, such an oligonucleotide is a gapmer. In certain such embodiments, the 3 '-wing of the gapmer may comprise any sugar modification motif.

In certain embodiments, the 5'- wing of a gapmer has a sugar motif selected from among those listed in the following non-limiting tables:

Table 2

Certain 5 '-Wing Sugar Motifs

Figure imgf000055_0001
AACCC ACCBA BBBCB CABAC CCACB

A BAA A ACCBB BBBCC CABBA CCACC

ABAAB ACCBC BBCAA CABBB CCBAA

ABAAC ACCCA BBCAB CABBC CCBAB

ABABA ACCCB BBCAC CABCA CCBAC

ABABB ACCCC BBCBA CABCB CCBBA

ABABC BAAAA BBCBB CABCC CCBBB

ABACA BAAAB BBCBC CACAA CCBBC

ABACB BAAAC BBCCA CACAB CCBCA

ABACC BAABA BBCCB CACAC CCBCB

ABBAA BAABB BBCCC CACBA CCBCC

ABBAB BAABC BCAAA CACBB CCCAA

ABBAC BAACA BCAAB CACBC CCCAB

ABBBA BAACB BCAAC CACCA CCCAC

ABBBB BAACC BCABA CACCB CCCBA

ABBBC BABAA BCABB CACCC CCCBB

ABBCA BABAB BCABC CBAAA CCCBC

ABBCB BABAC BCACA CBAAB CCCCA

ABBCC BABBA BCACB CBAAC CCCCB

ABCAA BABBB BCACC CBABA CCCCC

ABCAB BABBC BCBAA CBABB

ABCAC BABCA BCBAB CBABC

ABCBA BABCB BCBAC CBACA

Table 3

Certain 5 '-Wing Sugar Motifs

Certain 5 '-Wing Sugar Motifs

AAAAA BABC CBAB ABBB BAA

AAAAB BACA CBAC BAAA BAB

AAABA BACB CBBA BAAB BBA

AAABB BACC CBBB BABA BBB

AABAA BBAA CBBC BABB AA

AABAB BBAB CBCA BBAA AB

AABBA BBAC CBCB BBAB AC

AABBB BBBA CBCC BBBA BA

A BAA A BBBB CCAA BBBB BB

ABAAB BBBC CCAB AAA BC

ABABA BBCA CCAC AAB CA

ABABB BBCB CCBA AAC CB

ABBAA BBCC CCBB ABA CC

ABBAB BCAA CCBC ABB AA

ABBBA BCAB CCCA ABC AB ABBBB BCAC CCCB ACA BA

BAAAA ABCB BCBA ACB

BAAAB ABCC BCBB ACC

BAABA ACAA BCBC BAA

BAABB ACAB BCCA BAB

BABAA ACAC BCCB BAC

BABAB ACBA BCCC BBA

BABBA ACBB CAAA BBB

BABBB ACBC CAAB BBC

BBAAA ACCA CAAC BCA

BBAAB ACCB CABA BCB

BBABA ACCC CABB BCC

BBABB BAAA CABC CAA

BBBAA BAAB CACA CAB

BBBAB BAAC CACB CAC

BBBBA BABA CACC CBA

BBBBB BABB CBAA CBB

AAAA AACC CCCC CBC

AAAB ABAA AAAA CCA

AAAC ABAB AAAB CCB

AABA ABAC AABA CCC

AABB ABBA AABB AAA

AABC ABBB ABAA AAB

AACA ABBC ABAB ABA

AACB ABCA ABBA ABB

In certain embodiments, each A, each B, and each C located at the 3 '-most 5 '-wing nucleoside is a modified nucleoside. For example, in certain embodiments the 5 '-wing motif is selected from among ABB, BBB, and CBB, wherein the underlined nucleoside represents the 3 '-most 5 '-wing nucleoside and wherein the underlined nucleoside is a modified nucleoside.

In certain embodiments, each A comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, each A comprises a modified sugar moiety. In certain embodiments, each A comprises a 2'- substituted sugar moiety. In certain embodiments, each A comprises a 2 '-substituted sugar moiety selected from among F, ara-F, OCH3 and 0(CH2)2-OCH3. In certain embodiments, each A comprises a bicyclic sugar moiety. In certain embodiments, each A comprises a bicyclic sugar moiety selected from among cEt, cMOE, LNA, a-L-LNA, ENA and 2'-thio LNA. In certain embodiments, each A comprises a modified nucleobase. In certain embodiments, each A comprises a modified nucleobase selected from among 2-thio-thymidine nucleoside and 5-propyne uridine nucleoside. In certain embodiments, each A comprises an UNA. In certain embodiments, each A comprises an F-HNA. In certain embodiments, each B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, each B comprises a modified sugar moiety. In certain embodiments, each B comprises a 2'- substituted sugar moiety. In certain embodiments, each B comprises a 2'-subsituted sugar moiety selected from among F, (ara)-F, OCH3 and 0(CH2)2-OCH3. In certain embodiments, each B comprises a bicyclic sugar moiety. In certain embodiments, each B comprises a bicyclic sugar moiety selected from among cEt, cMOE, LNA, a-L-LNA, ENA and 2'-thio LNA. In certain embodiments, each B comprises a modified nucleobase. In certain embodiments, each B comprises a modified nucleobase selected from among 2-thio- thymidine nucleoside and 5-propyne urindine nucleoside. In certain embodiments, each B comprises an HNA. In certain embodiments, each B comprises an F-HNA.

In certain embodiments, each C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, each C comprises a modified sugar moiety. In certain embodiments, each C comprises a 2'- substituted sugar moiety. In certain embodiments, each C comprises a 2 '-substituted sugar moiety selected from among F, (ara)-F, OCH3 and 0(CH2)2-OCH3. In certain embodiments, each C comprises a 5'- substituted sugar moiety. In certain embodiments, each C comprises a 5 '-substituted sugar moiety selected from among 5 '-Me, and 5'-(R)-Me. In certain embodiments, each C comprises a bicyclic sugar moiety. In certain embodiments, each C comprises a bicyclic sugar moiety selected from among cEt, cMOE, LNA, a-L- LNA, ENA and 2'-thio LNA. In certain embodiments, each C comprises a modified nucleobase. In certain embodiments, each C comprises a modified nucleobase selected from among 2-thio-thymidine and 5-propyne uridine. In certain embodiments, each C comprises a 2-thio-thymidine nucleoside. In certain embodiments, each C comprises an UNA. In certain embodiments, each C comprises an F-HNA.

In certain embodiments, at least one of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety, and the other comprises a 2'-substituted sugar moiety. In certain embodiments, at least one of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety, and the other comprises a bicyclic sugar moiety.

In certain embodiments, at least one of A or B comprises a bicyclic sugar moiety, and the other comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-substituted sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside and the other of A or B comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-MOE sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2'- MOE sugar moiety. In certain embodiments, one of A or B is an a -L-LNA nucleoside and the other of A or B comprises a 2'-MOE sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is an a-L- LNA nucleoside and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside and the other of A or B comprises a 2'- (ara)-F sugar moiety.

In certain embodiments, at least one of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety, and the other comprises a 2'-substituted sugar moiety. In certain embodiments, one of A or B is an unmodified 2'-deoxyfuranose sugar moiety and the other of A or B comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an unmodified 2'-deoxyfuranose sugar moiety and the other of A or B comprises a 2'-MOE sugar moiety. In certain embodiments, one of A or B is an unmodified 2'- deoxyfuranose sugar moiety and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is an unmodified 2'-deoxyfuranose sugar moiety and the other of A or B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, at least one of A or B comprises a bicyclic sugar moiety, and the other comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises an unmodified 2'- deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises an unmodified 2'- deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an a -L-LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2 '-substituted sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2 '-substituted sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2 '-substituted sugar moiety. In certain embodiments, A is an a-L-LNA nucleoside and B comprises a 2 '-substituted sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2'-MOE sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2'-MOE sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2'-MOE sugar moiety. In certain

embodiments, A is an a-L-LNA nucleoside and B comprises a 2'-MOE sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2'-F sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2'-F sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2'-F sugar moiety. In certain embodiments, A is an a- L-LNA nucleoside and B comprises a 2'-F sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, A is an a-L-LNA nucleoside and B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, B comprises a bicyclic sugar moiety, and A comprises a 2'-MOE sugar moiety. In certain embodiments, B is an LNA nucleoside and A comprises a 2'-MOE sugar moiety. In certain embodiments, B is a cEt nucleoside and A comprises a 2'-MOE sugar moiety. In certain

embodiments, B is an a-L-LNA nucleoside and A comprises a 2'-MOE sugar moiety.

In certain embodiments, B comprises a bicyclic sugar moiety, A comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, B is an LNA nucleoside, A comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, B is a cEt nucleoside, A comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, B is an a-L-LNA nucleoside and A comprises a 2'-MOE sugar moiety.

In certain embodiments, B comprises a bicyclic sugar moiety, and A comprises a 2'-F sugar moiety. In certain embodiments, B is an LNA nucleoside and A comprises a 2'-F sugar moiety. In certain embodiments, B is a cEt nucleoside and A comprises a 2'-F sugar moiety. In certain embodiments, B is an a- L-LNA nucleoside and A comprises a 2'-F sugar moiety.

In certain embodiments, B comprises a bicyclic sugar moiety, and A comprises a 2'-(ara)-F sugar moiety. In certain embodiments, B is an LNA nucleoside and A comprises a 2'-(ara)-F sugar moiety. In certain embodiments, B is a cEt nucleoside and A comprises a 2'-(ara)-F sugar moiety. In certain embodiments, B is an a-L-LNA nucleoside and A comprises a 2'-(ara)-F sugar moiety.

In certain embodiments, at least one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2 '-substituted sugar moiety and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and Ccomprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and Ccomprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and Ccomprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2-thio- thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2- thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain

embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises 2-thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain

embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain

embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)- F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'- MOE sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an a- L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an a-L- LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar HNA surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F- HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F-HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'- F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an a-L- LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a F-HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-Me DNA sugar moiety. In certain embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain

embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^)-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain

embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^)-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^)-Me DNA sugar moiety.

In certain embodiments, at least two of A, B or C comprises a 2 '-substituted sugar moiety, and the other comprises a bicyclic sugar moiety. In certain embodiments, at least two of A, B or C comprises a bicyclic sugar moiety, and the other comprises a 2 '-substituted sugar moiety.

In certain embodiments, at least two of A, B or C comprises a 2 '-substituted sugar moiety, and the other comprises an unmodified 2'-deoxyfuranose sugar moiety.. In certain embodiments, at least two of A, B or C comprises a bicyclic sugar moiety, and the other comprises an unmodified 2'-deoxyfuranose sugar moiety.

Certain 3 '-wings

In certain embodiments, the 3'- wing of a gapmer consists of 1 to 5 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 2 to 5 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 3 to 5 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 4 or 5 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 1 to 4 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 1 to 3 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 1 or 2 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 2 to 4 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 2 or 3 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 3 or 4 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 1 nucleoside. In certain embodiments, the 3'- wing of a gapmer consists of 2 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 31inked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 4 linked nucleosides. In certain embodiments, the 3'- wing of a gapmer consists of 5 linked nucleosides.

In certain embodiments, the 3'- wing of a gapmer comprises at least one bicyclic nucleoside. In certain embodiments, the 3'- wing of a gapmer comprises at least one constrained ethyl nucleoside. In certain embodiments, the 3'- wing of a gapmer comprises at least one LNA nucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a bicyclic nucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a constrained ethyl nucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a LNA nucleoside.

In certain embodiments, the 3'- wing of a gapmer comprises at least one non-bicyclic modified nucleoside. In certain embodiments, the 3'- wing of a gapmer comprises at least two non-bicyclic modified nucleosides. In certain embodiments, the 3'- wing of a gapmer comprises at least three non-bicyclic modified nucleosides. In certain embodiments, the 3'- wing of a gapmer comprises at least four non-bicyclic modified nucleosides. In certain embodiments, the 3'- wing of a gapmer comprises at least one 2 '-substituted nucleoside. In certain embodiments, the 3'- wing of a gapmer comprises at least one 2'-MOE nucleoside. In certain embodiments, the 3'- wing of a gapmer comprises at least one 2'-OMe nucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a non-bicyclic modified nucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a 2 '-substituted nucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a 2'-MOE nucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a 2'-OMe nucleoside.

In certain embodiments, the 3'- wing of a gapmer comprises at least one 2'-deoxynucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a 2'-deoxynucleoside. In a certain embodiments, the 3'- wing of a gapmer comprises at least one ribonucleoside. In certain embodiments, each nucleoside of the 3'- wing of a gapmer is a ribonucleoside.

In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one non-bicyclic modified nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2 '-substituted nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2'-MOE nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2'-OMe nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one non-bicyclic modified nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2 '-substituted nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2'-MOE nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2'-OMe nucleoside. In certain embodiments, the 3'-wing of a gapmer comprises at least one constrained ethyl nucleoside and at least one 2'-deoxynucleoside.

In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside and at least one non-bicyclic modified nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'-substituted nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'-MOE nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'-OMe nucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside and at least one 2'- deoxynucleoside.

In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one non-bicyclic modified nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3'- wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one non-bicyclic modified nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside, at least one non-bicyclic modified nucleoside, and at least one 2'- deoxynucleoside.

In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one 2 '-substituted nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one 2'-substituted nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside, at least one 2 '-substituted nucleoside, and at least one 2'-deoxynucleoside.

In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one 2'-MOE nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one 2'-MOE nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside, at least one 2'-MOE nucleoside, and at least one 2'-deoxynucleoside.

In certain embodiments, the 3 '-wing of a gapmer comprises at least one bicyclic nucleoside, at least one 2'-OMe nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one constrained ethyl nucleoside, at least one 2'-OMe nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3 '-wing of a gapmer comprises at least one LNA nucleoside, at least one 2'-OMe nucleoside, and at least one 2'-deoxynucleoside. In certain embodiments, the 3'- wing of a gapmer has a sugar motif selected from among those listed the following non-limiting table:

Table 4

Certain 3 '-Wing Sugar Motifs

Figure imgf000068_0001
7j L-D-K-A 7k K-D-L-A 71 K-D-L-E

8j B-K-L-B 8k K-L-L-L 81 K-K-L-K

¾ K-D-D-B 9k K-D-D-L 91 L-D-D-K

10j A-K-B-A 10k A-K-L-A 101 A-B-K-A

lm E-E

In the above table, "A" represents a nucleoside comprising a 2'-substituted sugar moiety; "B" represents a bicyclic nucleoside; "D" represents a 2'-deoxynucleoside; "K" represents a constrained ethyl nucleoside; "L" represents an LNA nucleoside; "E" represents a 2'-MOE nucleoside; and "M" represents a 2'-OMe nucleoside.

In certain embodiments, an oligonucleotide comprises any 3 '-wing motif provided herein. In certain such embodiments, the oligonucleotide is a 3'-hemimer (does not comprise a 5 '-wing). In certain embodiments, such an oligonucleotide is a gapmer. In certain such embodiments, the 5 '-wing of the gapmer may comprise any sugar modification motif.

In certain embodiments, the 5'- wing of a gapmer has a sugar motif selected from among those listed in the following non-limiting tables:

Table 5

Certain 3 '-Wing Sugar Motifs

Figure imgf000069_0001
AACBA ACBCB BBBAC CAACA CCAAC

AACBB ACBCC BBBBA CAACB CCABA

AACBC ACCAA BBBBB CAACC CCABB

AACCA ACCAB BBBBC CABAA CCABC

AACCB ACCAC BBBC A CABAB CCACA

AACCC ACCBA BBBCB CABAC CCACB

A BAA A ACCBB BBBCC CABBA CCACC

ABAAB ACCBC BBCAA CABBB CCBAA

ABAAC ACCCA BBCAB CABBC CCBAB

ABABA ACCCB BBCAC CABCA CCBAC

ABABB ACCCC BBCBA CABCB CCBBA

ABABC BAAAA BBCBB CABCC CCBBB

ABACA BAAAB BBCBC CACAA CCBBC

ABACB BAAAC BBCCA CACAB CCBCA

ABACC BAABA BBCCB CACAC CCBCB

ABBAA BAABB BBCCC CACBA CCBCC

ABBAB BAABC BCAAA CACBB CCCAA

ABBAC BAACA BCAAB CACBC CCCAB

ABBBA BAACB BCAAC CACCA CCCAC

ABBBB BAACC BCABA CACCB CCCBA

ABBBC BABAA BCABB CACCC CCCBB

ABBCA BABAB BCABC CBAAA CCCBC

ABBCB BABAC BCACA CBAAB CCCCA

ABBCC BABBA BCACB CBAAC CCCCB

ABCAA BABBB BCACC CBABA CCCCC

ABCAB BABBC BCBAA CBABB

ABCAC BABCA BCBAB CBABC

ABCBA BABCB BCBAC CBACA

Table 6

Certain 3 '-Wing Sugar Motifs

Certain 3 '-Wing Sugar Motifs

AAAAA BABC CBAB ABBB BAA

AAAAB BACA CBAC BAAA BAB

AAABA BACB CBBA BAAB BBA

AAABB BACC CBBB BABA BBB

AABAA BBAA CBBC BABB AA

AABAB BBAB CBCA BBAA AB

AABBA BBAC CBCB BBAB AC

AABBB BBBA CBCC BBBA BA

A BAA A BBBB CCAA BBBB BB

ABAAB BBBC CCAB AAA BC ABABA BBCA CCAC AAB CA

ABABB BBCB CCBA AAC CB

ABBAA BBCC CCBB ABA CC

ABBAB BCAA CCBC ABB AA

ABBBA BCAB CCCA ABC AB

ABBBB BCAC CCCB ACA BA

BAAAA ABCB BCBA ACB

BAAAB ABCC BCBB ACC

BAABA ACAA BCBC BAA

BAABB ACAB BCCA BAB

BABAA ACAC BCCB BAC

BABAB ACBA BCCC BBA

BABBA ACBB CAAA BBB

BABBB ACBC CAAB BBC

BBAAA ACCA CAAC BCA

BBAAB ACCB CABA BCB

BBABA ACCC CABB BCC

BBABB BAAA CABC CAA

BBBAA BAAB CACA CAB

BBBAB BAAC CACB CAC

BBBBA BABA CACC CBA

BBBBB BABB CBAA CBB

AAAA AACC CCCC CBC

AAAB ABAA AAAA CCA

AAAC ABAB AAAB CCB

AABA ABAC AABA CCC

AABB ABBA AABB AAA

AABC ABBB ABAA AAB

AACA ABBC ABAB ABA

AACB ABCA ABBA ABB

In certain embodiments, each A, each B, and each C located at the 5 '-most 3 '-wing region nucleoside is a modified nucleoside. For example, in certain embodiments the 3 '-wing motif is selected from among ABB, BBB, and CBB, wherein the underlined nucleoside represents the the 5'-most 3'-wing region nucleoside and wherein the underlined nucleoside is a modified nucleoside.

In certain embodiments, each A comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, each A comprises a modified sugar moiety. In certain embodiments, each A comprises a 2'- substituted sugar moiety. In certain embodiments, each A comprises a 2 '-substituted sugar moiety selected from among F, ara-F, OCH3 and 0(CH2)2-OCH3. In certain embodiments, each A comprises a bicyclic sugar moiety. In certain embodiments, each A comprises a bicyclic sugar moiety selected from among cEt, cMOE, LNA, α-L-LNA, ENA and 2'-thio LNA. In certain embodiments, each A comprises a modified nucleobase. In certain embodiments, each A comprises a modified nucleobase selected from among 2-thio-thymidine nucleoside and 5-propyne uridine nucleoside.

In certain embodiments, each B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, each B comprises a modified sugar moiety. In certain embodiments, each B comprises a 2'- substituted sugar moiety. In certain embodiments, each B comprises a 2'-subsituted sugar moiety selected from among F, (ara)-F, OCH3 and 0(CH2)2-OCH3. In certain embodiments, each B comprises a bicyclic sugar moiety. In certain embodiments, each B comprises a bicyclic sugar moiety selected from among cEt, cMOE, LNA, a-L-LNA, ENA and 2'-thio LNA. In certain embodiments, each B comprises a modified nucleobase. In certain embodiments, each B comprises a modified nucleobase selected from among 2-thio- thymidine nucleoside and 5-propyne urindine nucleoside.

In certain embodiments, each C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, each C comprises a modified sugar moiety. In certain embodiments, each C comprises a 2'- substituted sugar moiety. In certain embodiments, each C comprises a 2 '-substituted sugar moiety selected from among F, (ara)-F, OCH3 and 0(CH2)2-OCH3. In certain embodiments, each C comprises a 5'- substituted sugar moiety. In certain embodiments, each C comprises a 5 '-substituted sugar moiety selected from among 5 '-Me, and 5'-(^-Με. In certain embodiments, each C comprises a bicyclic sugar moiety. In certain embodiments, each C comprises a bicyclic sugar moiety selected from among cEt, cMOE, LNA, a-L- LNA, ENA and 2'-thio LNA. In certain embodiments, each C comprises a modified nucleobase. In certain embodiments, each C comprises a modified nucleobase selected from among 2-thio-thymidine and 5-propyne uridine. In certain embodiments, each C comprises a 2-thio-thymidine nucleoside.

In certain embodiments, at least one of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety, and the other comprises a 2'-substituted sugar moiety. In certain embodiments, at least one of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety, and the other comprises a bicyclic sugar moiety.

In certain embodiments, at least one of A or B comprises a bicyclic sugar moiety, and the other comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-substituted sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside and the other of A or B comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-MOE sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2'- MOE sugar moiety. In certain embodiments, one of A or B is an a -L-LNA nucleoside and the other of A or B comprises a 2'-MOE sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is an a-L- LNA nucleoside and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside and the other of A or B comprises a 2'- (ara)-F sugar moiety.

In certain embodiments, at least one of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety, and the other comprises a 2'-substituted sugar moiety. In certain embodiments, one of A or B is an unmodified 2'-deoxyfuranose sugar moiety and the other of A or B comprises a 2 '-substituted sugar moiety. In certain embodiments, one of A or B is an unmodified 2'-deoxyfuranose sugar moiety and the other of A or B comprises a 2'-MOE sugar moiety. In certain embodiments, one of A or B is an unmodified 2'- deoxyfuranose sugar moiety and the other of A or B comprises a 2'-F sugar moiety. In certain embodiments, one of A or B is an unmodified 2'-deoxyfuranose sugar moiety and the other of A or B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, at least one of A or B comprises a bicyclic sugar moiety, and the other comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises an unmodified 2'- deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside and the other of A or B comprises an unmodified 2'- deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an a -L-LNA nucleoside and the other of A or B comprises an unmodified 2'-deoxyfuranose sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2 '-substituted sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2 '-substituted sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2 '-substituted sugar moiety. In certain embodiments, A is an a-L-LNA nucleoside and B comprises a 2 '-substituted sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2'-MOE sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2'-MOE sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2'-MOE sugar moiety. In certain

embodiments, A is an a-L-LNA nucleoside and B comprises a 2'-MOE sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2'-F sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2'-F sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2'-F sugar moiety. In certain embodiments, A is an a- L-LNA nucleoside and B comprises a 2'-F sugar moiety.

In certain embodiments, A comprises a bicyclic sugar moiety, and B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, A is an LNA nucleoside and B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, A is a cEt nucleoside and B comprises a 2'-(ara)-F sugar moiety. In certain embodiments, A is an a-L-LNA nucleoside and B comprises a 2'-(ara)-F sugar moiety.

In certain embodiments, B comprises a bicyclic sugar moiety, and A comprises a 2'-MOE sugar moiety. In certain embodiments, B is an LNA nucleoside and A comprises a 2'-MOE sugar moiety. In certain embodiments, B is a cEt nucleoside and A comprises a 2'-MOE sugar moiety. In certain

embodiments, B is an a-L-LNA nucleoside and A comprises a 2'-MOE sugar moiety.

In certain embodiments, B comprises a bicyclic sugar moiety, A comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, B is an LNA nucleoside, A comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, B is a cEt nucleoside, A comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, B is an a-L-LNA nucleoside and A comprises a 2'-MOE sugar moiety.

In certain embodiments, B comprises a bicyclic sugar moiety, and A comprises a 2'-F sugar moiety. In certain embodiments, B is an LNA nucleoside and A comprises a 2'-F sugar moiety. In certain embodiments, B is a cEt nucleoside and A comprises a 2'-F sugar moiety. In certain embodiments, B is an a- L-LNA nucleoside and A comprises a 2'-F sugar moiety.

In certain embodiments, B comprises a bicyclic sugar moiety, and A comprises a 2'-(ara)-F sugar moiety. In certain embodiments, B is an LNA nucleoside and A comprises a 2'-(ara)-F sugar moiety. In certain embodiments, B is a cEt nucleoside and A comprises a 2'-(ara)-F sugar moiety. In certain embodiments, B is an a-L-LNA nucleoside and A comprises a 2'-(ara)-F sugar moiety.

In certain embodiments, at least one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2 '-substituted sugar moiety and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and Ccomprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and Ccomprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and Ccomprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a modified nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2 '-substituted sugar moiety, and C comprises a 2-thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 2-thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2-thio- thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 2- thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain

embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 2-thio-thymidine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises 2-thio-thymidine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain

embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises an unmodified 2'-deoxyfuranose sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain

embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5-propyne uridine nucleobase.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)- F sugar moiety, and C comprises a sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'- MOE sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an a- L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an a-L- LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a sugar HNA surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F- HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a F-HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'- F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an a-L- LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a F-HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a F-HNA sugar surrogate.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5 '-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-Me DNA sugar moiety. In certain embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5 '-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain

embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-MOE sugar moiety, and C comprises a 5'-(^)-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain

embodiments,one of A or B is an LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-F sugar moiety, and C comprises a 5'-(^)-Me DNA sugar moiety.

In certain embodiments, one of A or B comprises a bicyclic sugar moiety, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is a cEt nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^-Με DNA sugar moiety. In certain embodiments, one of A or B is an a-L-LNA nucleoside, another of A or B comprises a 2'-(ara)-F sugar moiety, and C comprises a 5'-(^)-Me DNA sugar moiety.

In certain embodiments, at least two of A, B or C comprises a 2 '-substituted sugar moiety, and the other comprises a bicyclic sugar moiety. In certain embodiments, at least two of A, B or C comprises a bicyclic sugar moiety, and the other comprises a 2 '-substituted sugar moiety.

In certain embodiments, at least two of A, B or C comprises a 2 '-substituted sugar moiety, and the other comprises an unmodified 2'-deoxyfuranose sugar moiety.. In certain embodiments, at least two of A, B or C comprises a bicyclic sugar moiety, and the other comprises an unmodified 2'-deoxyfuranose sugar moiety.

Certain gaps

In certain embodiments, the gap of a gapmer consists of 6 to 20 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 6 to 15 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 6 to 12 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 6 to 10 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 6 to 9 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 6 to 8 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 6 or 7 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 7 to 10 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 7 to 9 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 7 or 8 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 8 to 10 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 8 or 9 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 6 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 7 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 8 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 9 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 10 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 1 1 linked nucleosides. In certain embodiments, the gap of a gapmer consists of 12 linked nucleosides.

In certain embodiments, each nucleotide of the gap of a gapmer is a 2'-deoxynucleoside. In certain embodiments, the gap comprises one or more modified nucleosides. In certain embodiments, each nucleotide of the gap of a gapmer is a 2'-deoxynucleoside or is a modified nucleoside that is "DNA-like." In such embodiments, "DNA-like" means that the nucleoside has similar characteristics to DNA, such that a duplex comprising the gapmer and an RNA molecule is capable of activating RNase H. For example, under certain conditions, 2'- fiuoro (arabino) nucleosides (also referred to as FAN A) have been shown to support RNase H activation, and thus is DNA-like. In certain embodiments, one or more nucleosides of the gap of a gapmer is not a 2'-deoxynucleoside and is not DNA-like. In certain such embodiments, the gapmer nonetheless supports RNase H activation (e.g., by virtue of the number or placement of the non-DNA nucleosides).

Certain gapmer motifs

In certain embodiments, a gapmer comprises a 5 '-wing, a gap, and a 3' wing, wherein the 5 '-wing, gap, and 3 ' wing are independently selected from among those discussed above. For example, in certain embodiments, a gapmer has a 5 '-wing selected from any of the 5 '-wing motifs in Tables 1, 2, and 3 above and a 3 '-wing selected from any of the 3 '-wing motifs in Tables, 4, 5, and 6. For example, in certain embodiments, a gapmer has a 5 '-wing, a gap, and a 3 '-wing having features selected from among those listed in the following non-limiting table:

Table 7

Certain Gapmer Sugar Motifs

Figure imgf000080_0001
At least one non-bicyclic All 2'-deoxynucleosides At least one cEt nucleoside modified nucleoside

At least one 2 '-substituted All 2'-deoxynucleosides At least one bicyclic nucleoside nucleoside

At least one 2 '-substituted All 2'-deoxynucleosides At least one LNA nucleoside nucleoside

At least one 2 '-substituted All 2'-deoxynucleosides At least one cEt nucleoside nucleoside

At least one 2'-MOE nucleoside All 2'-deoxynucleosides At least one bicyclic

nucleoside

At least one 2'-MOE nucleoside All 2'-deoxynucleosides At least one LNA nucleoside

At least one 2'-MOE nucleoside All 2'-deoxynucleosides At least one cEt nucleoside

At least one 2'-OMe nucleoside All 2'-deoxynucleosides At least one bicyclic

nucleoside

At least one 2'-OMe nucleoside All 2'-deoxynucleosides At least one LNA nucleoside

At least one 2'-OMe nucleoside All 2'-deoxynucleosides At least one cEt nucleoside

At least one 2'-deoxynucleoside All 2'-deoxynucleosides At least one bicyclic

nucleoside

At least one 2'-deoxynucleoside All 2'-deoxynucleosides At least one LNA nucleoside

At least one 2'-deoxynucleoside All 2'-deoxynucleosides At least one cEt nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least one non-bicyclic modified nucleoside

At least one LNA nucleoside All 2'-deoxynucleosides At least one non-bicyclic modified nucleoside

At least one cEt nucleoside All 2'-deoxynucleosides At least one non-bicyclic modified nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least one 2'-substituted nucleoside

At least one LNA nucleoside All 2'-deoxynucleosides At least one 2'-substituted nucleoside

At least one cEt nucleoside All 2'-deoxynucleosides At least one 2'-substituted nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least one 2'-MOE

nucleoside

At least one LNA nucleoside All 2'-deoxynucleosides At least one 2'-MOE

nucleoside

At least one cEt nucleoside All 2'-deoxynucleosides At least one 2'-MOE

nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least one 2'-OMe

nucleoside

At least one LNA nucleoside All 2'-deoxynucleosides At least one 2'-OMe

nucleoside

At least one cEt nucleoside All 2'-deoxynucleosides At least one 2'-OMe

nucleoside At least one bicyclic nucleoside All 2'-deoxynucleosides At least one 2'- deoxynucleoside

At least one LNA nucleoside All 2'-deoxynucleosides At least one 2'- deoxynucleoside

At least one cEt nucleoside All 2'-deoxynucleosides At least one 2'- deoxynucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least one bicyclic and at least one 2 '-substituted nucleoside and at least one 2'- nucleoside substituted nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least two bicyclic and at least one 2 '-substituted nucleosides

nucleoside

At least one cEt nucleoside and All 2'-deoxynucleosides At least one bicyclic at least one 2'-substituted nucleoside and at least one 2'- nucleoside substituted nucleoside

At least one cEt nucleoside and All 2'-deoxynucleosides At least two bicyclic at least one 2'-substituted nucleosides

nucleoside

At least one LNA nucleoside and All 2'-deoxynucleosides At least one bicyclic at least one 2'-substituted nucleoside and at least one 2'- nucleoside substituted nucleoside

At least one LNA nucleoside and All 2'-deoxynucleosides At least two bicyclic at least one 2'-substituted nucleosides

nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least one LNA nucleoside and at least one 2 '-substituted and at least one 2 '-substituted nucleoside nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least two LNA nucleosides and at least one 2 '-substituted

nucleoside

At least one cEt nucleoside and All 2'-deoxynucleosides At least one LNA nucleoside at least one 2'-substituted and at least one 2 '-substituted nucleoside nucleoside

At least one cEt nucleoside and All 2'-deoxynucleosides At least two LNA nucleosides at least one 2'-substituted

nucleoside

At least one LNA nucleoside and All 2'-deoxynucleosides At least one LNA nucleoside at least one 2'-substituted and at least one 2 '-substituted nucleoside nucleoside

At least one LNA nucleoside and All 2'-deoxynucleosides At least two LNA nucleosides at least one 2'-substituted

nucleoside

At least one bicyclic nucleoside All 2'-deoxynucleosides At least one bicyclic and at least one 2'- nucleoside and at least one 2'- deoxynucleoside substituted nucleoside 44 At least one bicyclic nucleoside All 2'-deoxynucleosides At least two bicyclic and at least one 2'- nucleosides

deoxynucleoside

45 At least one cEt nucleoside and All 2'-deoxynucleosides At least one bicyclic

at least one 2'-deoxynucleoside nucleoside and at least one 2'- substituted nucleoside

46 At least one cEt nucleoside and All 2'-deoxynucleosides At least two bicyclic

at least one 2'-deoxynucleoside nucleosides

47 At least one LNA nucleoside and All 2'-deoxynucleosides At least one bicyclic

at least one 2'-deoxynucleoside nucleoside and at least one 2'- substituted nucleoside

48 At least one LNA nucleoside and All 2'-deoxynucleosides At least two bicyclic

at least one 2'-deoxynucleoside nucleosides

49 At least one bicyclic nucleoside All 2'-deoxynucleosides At least one LNA nucleoside and at least one 2'- and at least one 2 '-substituted deoxynucleoside nucleoside

50 At least one bicyclic nucleoside All 2'-deoxynucleosides At least two LNA nucleosides and at least one 2'- deoxynucleoside

51 At least one cEt nucleoside and All 2'-deoxynucleosides At least one LNA nucleoside at least one 2'-deoxynucleoside and at least one 2 '-substituted nucleoside

52 At least one cEt nucleoside and All 2'-deoxynucleosides At least two LNA nucleosides at least one 2'-deoxynucleoside

53 At least one LNA nucleoside and All 2'-deoxynucleosides At least one LNA nucleoside at least one 2'-deoxynucleoside and at least one 2 '-substituted nucleoside

54 At least one LNA nucleoside and All 2'-deoxynucleosides At least two LNA nucleosides at least one 2'-deoxynucleoside

55 At least two 2 '-substituted All 2'-deoxynucleosides At least one bicyclic

nucleosides nucleoside and at least one 2'- substituted nucleoside

56 At least two 2 '-substituted All 2'-deoxynucleosides At least two bicyclic

nucleosides nucleosides

57 At least two 2 '-substituted All 2'-deoxynucleosides At least one LNA nucleoside nucleosides and at least one 2 '-substituted nucleoside

58 At least two 2 '-substituted All 2'-deoxynucleosides At least two LNA nucleosides nucleosides

In certain embodiments, a gapmer comprises a 5 '-wing, a gap, and a 3' wing, wherein the 5 '-wing, gap, and 3 ' wing are independently selected from among those discussed above. For example, in certain embodiments, a gapmer has a 5 '-wing, a gap, and a 3 '-wing wherein the 5 '-wing and the 3 '-wing have features selected from among those listed in the tables above. In certain embodiments, any 5 '-wing may be paired with any 3 '-wing. In certain embodiments the 5 '-wing may comprise ABBBB and the 3 '-wing may comprise BBA. In certain embodiments the 5 '-wing may comprise ACACA and the 3 '-wing may comprise BB. For example, in certain embodiments, a gapmer has a 5 '-wing, a gap, and a 3 '-wing having features selected from among those listed in the following non-limiting table, wherein each motif is represented as (5'-wing)-(gap)-(3'-wing), wherein each number represents the number of linked nucleosides in each portion of the motif, for example, a 5- 10-5 motif would have a 5'-wing comprising 5 nucleosides, a gap comprising 10 nucleosides, and a 3 '-wing comprising 5 nucleosides:

Table 8

Certain Gapmer Sugar Motifs

Figure imgf000084_0001

In certain embodiments, gapmers have a motif described by Formula I as follows:

(A)m-(B)n-(J)p-(B)r-(J)t-(D)g.h-(J)v-(B)w-(J)x-(B)y-(A)z

wherein:

each A is independently a 2 '-substituted nucleoside;

each B is independently a bicyclic nucleoside;

each J is independently either a 2 '-substituted nucleoside or a 2'-deoxynucleoside;

each D is a 2'-deoxynucleoside;

m is 0-4; n is 0-2; p is 0-2; r is 0-2; t is 0-2; v is 0-2; w is 0-4; x is 0-2; y is 0-2; z is 0-4; g is 6; and h is 14;

provided that:

at least one of m, n, and r is other than 0;

at least one of w and y is other than 0; the sum of m, n, p, r, and t is from 2 to 5; and

the sum of v, w, x, y, and z is from 2 to 5.

In certain embodiments, one or more 2 '-substituted nucleoside is a 2'-MOE nucleoside. In certain embodiments, one or more 2'-substituted nucleoside is a 2'-OMe nucleoside. In certain In certain

embodiments, one or more bicyclic nucleoside is a cEt nucleoside. In certain embodiments, one or more bicyclic nucleoside is an LNA nucleoside.

In certain embodiments, a gapmer of Formula I has a motif selected from among gapmer motifs 1-58.

In certain embodiments, gapmers have a motif described by Formula II as follows:

(J)m-(B)n-(J)p-(B)r-(A)t-(D)g-(A)v-(B)w-(J)x-(B)y-(J)z

wherein:

each A is independently a 2 '-substituted nucleoside;

each B is independently a bicyclic nucleoside;

each J is independently either a 2 '-substituted nucleoside or a 2'-deoxynucleoside;

each D is a 2'-deoxynucleoside;

m is 0-4; n is 0-2; p is 0-2; r is 0-2; t is 0-2; v is 0-2; w is 0-4; x is 0-2; y is 0-2; z is 0-4; g is 6-14; provided that:

at least one of m, n, and r is other than 0;

at least one of w and y is other than 0;

the sum of m, n, p, r, and t is from 1 to 5; and

the sum of v, w, x, y, and z is from 1 to 5.

In certain embodiments, one or more 2 '-substituted nucleoside is a 2'-MOE nucleoside. In certain embodiments, one or more 2'-substituted nucleoside is a 2'-OMe nucleoside. In certain embodiments, one or more bicyclic nucleoside is a cEt nucleoside. In certain embodiments, one or more bicyclic nucleoside is an LNA nucleoside.

In certain embodiments, each 2 '-substituted nucleoside is a 2'-MOE nucleoside. In certain embodiments, each 2 '-substituted nucleoside is a 2'-OMe nucleoside. In certain embodiments, each bicyclic nucleoside is a cEt nucleoside. In certain embodiments, each bicyclic nucleoside is an LNA nucleoside.

In certain embodiments, each A is the same 2'-substituted nucleoside. In certain embodiments, each B is the same bicyclic nucleoside. In certain embodiments each A is the same 2 '-modified nucleoside and each B is the same bicyclic nucleoside. In certain embodiments, each J is a 2 '-modified nucleoside. In certain embodiments each J is the same 2 '-modified nucleoside. In certain embodiments, each J and each A is the same 2'-modified nucleoside.

In certain embodiments, a gapmer of Formula II has a motif selected from among gapmer motifs 1-

58.

In certain embodiments, a gapmer comprises a 5 '-wing, a gap, and a 3' wing, independently selected from among those proved in the above tables, for example as provided in the following table: Table 9

Certain Gapmer Sugar Motifs

Figure imgf000086_0001
98 18(a-i) All 2'-deoxynucleosides 2(a-i)

99 19(a-i) All 2'-deoxynucleosides 2(a-i)

100 20(a-i) All 2'-deoxynucleosides 2(a-i)

101 21(a-i) All 2'-deoxynucleosides 2(a-i)

102 22(a-i) All 2'-deoxynucleosides 2(a-i)

103 l(a-i) All 2'-deoxynucleosides 3(a-i)

104 2(a-i) All 2'-deoxynucleosides 3(a-i)

105 3(a-i) All 2'-deoxynucleosides 3(a-i)

106 4(a-i) All 2'-deoxynucleosides 3(a-i)

107 5(a-i) All 2'-deoxynucleosides 3(a-i)

108 6(a-i) All 2'-deoxynucleosides 3(a-i)

109 7(a-i) All 2'-deoxynucleosides 3(a-i)

1 10 8(a-i) All 2'-deoxynucleosides 3(a-i)

1 1 1 9(a-i) All 2'-deoxynucleosides 3(a-i)

1 12 10(a-i) All 2'-deoxynucleosides 3(a-i)

1 13 l l(a-i) All 2'-deoxynucleosides 3(a-i)

1 14 12(a-i) All 2'-deoxynucleosides 3(a-i)

1 15 13(a-i) All 2'-deoxynucleosides 3(a-i)

1 16 14(a-i) All 2'-deoxynucleosides 3(a-i)

1 17 15(a-i) All 2'-deoxynucleosides 3(a-i)

1 18 16(a-i) All 2'-deoxynucleosides 3(a-i)

1 19 17(a-i) All 2'-deoxynucleosides 3(a-i)

120 18(a-i) All 2'-deoxynucleosides 3(a-i)

121 19(a-i) All 2'-deoxynucleosides 3(a-i)

122 20(a-i) All 2'-deoxynucleosides 3(a-i)

123 21(a-i) All 2'-deoxynucleosides 3(a-i)

124 22(a-i) All 2'-deoxynucleosides 3(a-i)

125 l(a-i) All 2'-deoxynucleosides 4(a-i)

126 2(a-i) All 2'-deoxynucleosides 4(a-i)

127 3(a-i) All 2'-deoxynucleosides 4(a-i)

128 4(a-i) All 2'-deoxynucleosides 4(a-i)

129 5(a-i) All 2'-deoxynucleosides 4(a-i)

130 6(a-i) All 2'-deoxynucleosides 4(a-i)

131 7(a-i) All 2'-deoxynucleosides 4(a-i)

132 8(a-i) All 2'-deoxynucleosides 4(a-i)

133 9(a-i) All 2'-deoxynucleosides 4(a-i)

134 10(a-i) All 2'-deoxynucleosides 4(a-i)

135 l l(a-i) All 2'-deoxynucleosides 4(a-i)

136 12(a-i) All 2'-deoxynucleosides 4(a-i)

137 13(a-i) All 2'-deoxynucleosides 4(a-i)

138 14(a-i) All 2'-deoxynucleosides 4(a-i)

139 15(a-i) All 2'-deoxynucleosides 4(a-i)

140 16(a-i) All 2'-deoxynucleosides 4(a-i)

141 17(a-i) All 2'-deoxynucleosides 4(a-i)

142 18(a-i) All 2'-deoxynucleosides 4(a-i) 143 19(a-i) All 2'-deoxynucleosides 4(a-i)

144 20(a-i) All 2'-deoxynucleosides 4(a-i)

145 21(a-i) All 2'-deoxynucleosides 4(a-i)

146 22(a-i) All 2'-deoxynucleosides 4(a-i)

147 l(a-i) All 2'-deoxynucleosides 5(a-i)

148 2(a-i) All 2'-deoxynucleosides 5(a-i)

149 3(a-i) All 2'-deoxynucleosides 5(a-i)

150 4(a-i) All 2'-deoxynucleosides 5(a-i)

151 5(a-i) All 2'-deoxynucleosides 5(a-i)

152 6(a-i) All 2'-deoxynucleosides 5(a-i)

153 7(a-i) All 2'-deoxynucleosides 5(a-i)

154 8(a-i) All 2'-deoxynucleosides 5(a-i)

155 9(a-i) All 2'-deoxynucleosides 5(a-i)

156 10(a-i) All 2'-deoxynucleosides 5(a-i)

157 l l(a-i) All 2'-deoxynucleosides 5(a-i)

158 12(a-i) All 2'-deoxynucleosides 5(a-i)

159 13(a-i) All 2'-deoxynucleosides 5(a-i)

160 14(a-i) All 2'-deoxynucleosides 5(a-i)

161 15(a-i) All 2'-deoxynucleosides 5(a-i)

162 16(a-i) All 2'-deoxynucleosides 5(a-i)

163 17(a-i) All 2'-deoxynucleosides 5(a-i)

164 18(a-i) All 2'-deoxynucleosides 5(a-i)

165 19(a-i) All 2'-deoxynucleosides 5(a-i)

166 20(a-i) All 2'-deoxynucleosides 5(a-i)

167 21(a-i) All 2'-deoxynucleosides 5(a-i)

168 22(a-i) All 2'-deoxynucleosides 5(a-i)

169 l(a-i) All 2'-deoxynucleosides 6(a-i)

170 2(a-i) All 2'-deoxynucleosides 6(a-i)

171 3(a-i) All 2'-deoxynucleosides 6(a-i)

172 4(a-i) All 2'-deoxynucleosides 6(a-i)

173 5(a-i) All 2'-deoxynucleosides 6(a-i)

174 6(a-i) All 2'-deoxynucleosides 6(a-i)

175 7(a-i) All 2'-deoxynucleosides 6(a-i)

176 8(a-i) All 2'-deoxynucleosides 6(a-i)

177 9(a-i) All 2'-deoxynucleosides 6(a-i)

178 10(a-i) All 2'-deoxynucleosides 6(a-i)

179 l l(a-i) All 2'-deoxynucleosides 6(a-i)

180 12(a-i) All 2'-deoxynucleosides 6(a-i)

181 13(a-i) All 2'-deoxynucleosides 6(a-i)

182 14(a-i) All 2'-deoxynucleosides 6(a-i)

183 15(a-i) All 2'-deoxynucleosides 6(a-i)

184 16(a-i) All 2'-deoxynucleosides 6(a-i)

184 17(a-i) All 2'-deoxynucleosides 6(a-i)

186 18(a-i) All 2'-deoxynucleosides 6(a-i)

187 19(a-i) All 2'-deoxynucleosides 6(a-i) 188 20(a-i) All 2'-deoxynucleosides 6(a-i)

189 21(a-i) All 2'-deoxynucleosides 6(a-i)

190 22(a-i) All 2'-deoxynucleosides 6(a-i)

191 l(a-i) All 2'-deoxynucleosides 7(a-i)

192 2(a-i) All 2'-deoxynucleosides 7(a-i)

193 3(a-i) All 2'-deoxynucleosides 7(a-i)

194 4(a-i) All 2'-deoxynucleosides 7(a-i)

195 5(a-i) All 2'-deoxynucleosides 7(a-i)

196 6(a-i) All 2'-deoxynucleosides 7(a-i)

197 7(a-i) All 2'-deoxynucleosides 7(a-i)

198 8(a-i) All 2'-deoxynucleosides 7(a-i)

199 9(a-i) All 2'-deoxynucleosides 7(a-i)

200 10(a-i) All 2'-deoxynucleosides 7(a-i)

201 l l(a-i) All 2'-deoxynucleosides 7(a-i)

202 12(a-i) All 2'-deoxynucleosides 7(a-i)

203 13(a-i) All 2'-deoxynucleosides 7(a-i)

204 14(a-i) All 2'-deoxynucleosides 7(a-i)

205 15(a-i) All 2'-deoxynucleosides 7(a-i)

206 16(a-i) All 2'-deoxynucleosides 7(a-i)

207 17(a-i) All 2'-deoxynucleosides 7(a-i)

208 18(a-i) All 2'-deoxynucleosides 7(a-i)

209 19(a-i) All 2'-deoxynucleosides 7(a-i)

210 20(a-i) All 2'-deoxynucleosides 7(a-i)

21 1 21(a-i) All 2'-deoxynucleosides 7(a-i)

212 22(a-i) All 2'-deoxynucleosides 7(a-i)

213 l(a-i) All 2'-deoxynucleosides 8(a-i)

214 2(a-i) All 2'-deoxynucleosides 8(a-i)

215 3(a-i) All 2'-deoxynucleosides 8(a-i)

216 4(a-i) All 2'-deoxynucleosides 8(a-i)

217 5(a-i) All 2'-deoxynucleosides 8(a-i)

218 6(a-i) All 2'-deoxynucleosides 8(a-i)

219 7(a-i) All 2'-deoxynucleosides 8(a-i)

220 8(a-i) All 2'-deoxynucleosides 8(a-i)

221 9(a-i) All 2'-deoxynucleosides 8(a-i)

222 10(a-i) All 2'-deoxynucleosides 8(a-i)

223 l l(a-i) All 2'-deoxynucleosides 8(a-i)

224 12(a-i) All 2'-deoxynucleosides 8(a-i)

225 13(a-i) All 2'-deoxynucleosides 8(a-i)

226 14(a-i) All 2'-deoxynucleosides 8(a-i)

227 15(a-i) All 2'-deoxynucleosides 8(a-i)

228 16(a-i) All 2'-deoxynucleosides 8(a-i)

229 17(a-i) All 2'-deoxynucleosides 8(a-i)

230 18(a-i) All 2'-deoxynucleosides 8(a-i)

231 19(a-i) All 2'-deoxynucleosides 8(a-i)

232 20(a-i) All 2'-deoxynucleosides 8(a-i) 233 21(a-i) All 2'-deoxynucleosides 8(a-i)

234 22(a-i) All 2'-deoxynucleosides 8(a-i)

235 l(a-i) All 2'-deoxynucleosides 9(a-i)

236 2(a-i) All 2'-deoxynucleosides 9(a-i)

237 3(a-i) All 2'-deoxynucleosides 9(a-i)

238 4(a-i) All 2'-deoxynucleosides 9(a-i)

239 5(a-i) All 2'-deoxynucleosides 9(a-i)

240 6(a-i) All 2'-deoxynucleosides 9(a-i)

241 7(a-i) All 2'-deoxynucleosides 9(a-i)

242 8(a-i) All 2'-deoxynucleosides 9(a-i)

243 9(a-i) All 2'-deoxynucleosides 9(a-i)

244 10(a-i) All 2'-deoxynucleosides 9(a-i)

245 l l(a-i) All 2'-deoxynucleosides 9(a-i)

246 12(a-i) All 2'-deoxynucleosides 9(a-i)

247 13(a-i) All 2'-deoxynucleosides 9(a-i)

248 14(a-i) All 2'-deoxynucleosides 9(a-i)

249 15(a-i) All 2'-deoxynucleosides 9(a-i)

250 16(a-i) All 2'-deoxynucleosides 9(a-i)

251 17(a-i) All 2'-deoxynucleosides 9(a-i)

252 18(a-i) All 2'-deoxynucleosides 9(a-i)

253 19(a-i) All 2'-deoxynucleosides 9(a-i)

254 20(a-i) All 2'-deoxynucleosides 9(a-i)

255 21(a-i) All 2'-deoxynucleosides 9(a-i)

256 22(a-i) All 2'-deoxynucleosides 9(a-i)

257 l(a-i) All 2'-deoxynucleosides 10(a-i)

258 2(a-i) All 2'-deoxynucleosides 10(a-i)

259 3(a-i) All 2'-deoxynucleosides 10(a-i)

260 4(a-i) All 2'-deoxynucleosides 10(a-i)

261 5(a-i) All 2'-deoxynucleosides 10(a-i)

262 6(a-i) All 2'-deoxynucleosides 10(a-i)

263 7(a-i) All 2'-deoxynucleosides 10(a-i)

264 8(a-i) All 2'-deoxynucleosides 10(a-i)

265 9(a-i) All 2'-deoxynucleosides 10(a-i)

266 10(a-i) All 2'-deoxynucleosides 10(a-i)

267 l l(a-i) All 2'-deoxynucleosides 10(a-i)

268 12(a-i) All 2'-deoxynucleosides 10(a-i)

269 13(a-i) All 2'-deoxynucleosides 10(a-i)

270 14(a-i) All 2'-deoxynucleosides 10(a-i)

271 15(a-i) All 2'-deoxynucleosides 10(a-i)

272 16(a-i) All 2'-deoxynucleosides 10(a-i)

273 17(a-i) All 2'-deoxynucleosides 10(a-i)

274 18(a-i) All 2'-deoxynucleosides 10(a-i)

275 19(a-i) All 2'-deoxynucleosides 10(a-i)

276 20(a-i) All 2'-deoxynucleosides 10(a-i)

277 21(a-i) All 2'-deoxynucleosides 10(a-i) 278 22(a-i) All 2'-deoxynucleosides 10(a-i)

279 l(a)-22(a) All 2'-deoxynucleosides 1(a)- 10(a)

280 l(b)-22(b) All 2'-deoxynucleosides 1(a)- 10(a)

281 l(c)-22(c) All 2'-deoxynucleosides 1(a)- 10(a)

282 l(d)-22(d) All 2'-deoxynucleosides 1(a)- 10(a)

283 l(e)-22(e) All 2'-deoxynucleosides 1(a)- 10(a)

284 l(f)-22(f) All 2'-deoxynucleosides 1(a)- 10(a)

285 l(g)-22(g) All 2'-deoxynucleosides 1(a)- 10(a)

286 l(h)-22(h) All 2'-deoxynucleosides 1(a)- 10(a)

287 l(i)-22(i) All 2'-deoxynucleosides 1(a)- 10(a)

288 l(a)-22(a) All 2'-deoxynucleosides 1(b)- 10(b)

289 l(b)-22(b) All 2'-deoxynucleosides 1(b)- 10(b)

290 l(c)-22(c) All 2'-deoxynucleosides 1(b)- 10(b)

291 l(d)-22(d) All 2'-deoxynucleosides 1(b)- 10(b)

292 l(e)-22(e) All 2'-deoxynucleosides 1(b)- 10(b)

293 l(f)-22(f) All 2'-deoxynucleosides 1(b)- 10(b)

294 l(g)-22(g) All 2'-deoxynucleosides 1(b)- 10(b)

295 l(h)-22(h) All 2'-deoxynucleosides 1(b)- 10(b)

296 l(i)-22(i) All 2'-deoxynucleosides 1(b)- 10(b)

297 l(a)-22(a) All 2'-deoxynucleosides 1(c)- 10(c)

298 l(b)-22(b) All 2'-deoxynucleosides 1(c)- 10(c)

299 l(c)-22(c) All 2'-deoxynucleosides 1(c)- 10(c)

300 l(d)-22(d) All 2'-deoxynucleosides 1(c)- 10(c)

301 l(e)-22(e) All 2'-deoxynucleosides 1(c)- 10(c)

302 l(f)-22(f) All 2'-deoxynucleosides 1(c)- 10(c)

303 l(g)-22(g) All 2'-deoxynucleosides 1(c)- 10(c)

304 l(h)-22(h) All 2'-deoxynucleosides 1(c)- 10(c)

305 l(i)-22(i) All 2'-deoxynucleosides 1(c)- 10(c)

306 l(a)-22(a) All 2'-deoxynucleosides 1(d)- 10(d)

307 l(b)-22(b) All 2'-deoxynucleosides 1(d)- 10(d)

308 l(c)-22(c) All 2'-deoxynucleosides 1(d)- 10(d)

309 l(d)-22(d) All 2'-deoxynucleosides 1(d)- 10(d)

310 l(e)-22(e) All 2'-deoxynucleosides 1(d)- 10(d)

31 1 l(f)-22(f) All 2'-deoxynucleosides 1(d)- 10(d)

312 l(g)-22(g) All 2'-deoxynucleosides 1(d)- 10(d)

313 l(h)-22(h) All 2'-deoxynucleosides 1(d)- 10(d)

314 l(i)-22(i) All 2'-deoxynucleosides 1(d)- 10(d)

315 l(a)-22(a) All 2'-deoxynucleosides 1(e)- 10(e)

316 l(b)-22(b) All 2'-deoxynucleosides 1(e)- 10(e)

317 l(c)-22(c) All 2'-deoxynucleosides 1(e)- 10(e)

318 l(d)-22(d) All 2'-deoxynucleosides 1(e)- 10(e)

319 l(e)-22(e) All 2'-deoxynucleosides 1(e)- 10(e)

320 l(f)-22(f) All 2'-deoxynucleosides 1(e)- 10(e)

321 l(g)-22(g) All 2'-deoxynucleosides 1(e)- 10(e)

322 l(h)-22(h) All 2'-deoxynucleosides 1(e)- 10(e) 323 l(i)-22(i) All 2'-deoxynucleosides l(e)-10(e)

324 l(a)-22(a) All 2'-deoxynucleosides l(f)-10(f)

325 l(b)-22(b) All 2'-deoxynucleosides l(f)-10(f)

326 l(c)-22(c) All 2'-deoxynucleosides l(f)-10(f)

327 l(d)-22(d) All 2'-deoxynucleosides l(f)-10(f)

328 l(e)-22(e) All 2'-deoxynucleosides l(f)-10(f)

329 l(f)-22(f) All 2'-deoxynucleosides l(f)-10(f)

330 l(g)-22(g) All 2'-deoxynucleosides l(f)-10(f)

331 l(h)-22(h) All 2'-deoxynucleosides l(f)-10(f)

332 l(i)-22(i) All 2'-deoxynucleosides l(f)-10(f)

333 l(a)-22(a) All 2'-deoxynucleosides l(g)-10(g)

334 l(b)-22(b) All 2'-deoxynucleosides l(g)-10(g)

335 l(c)-22(c) All 2'-deoxynucleosides l(g)-10(g)

336 l(d)-22(d) All 2'-deoxynucleosides l(g)-10(g)

337 l(e)-22(e) All 2'-deoxynucleosides l(g)-10(g)

338 l(f)-22(f) All 2'-deoxynucleosides l(g)-10(g)

339 l(g)-22(g) All 2'-deoxynucleosides l(g)-10(g)

340 l(h)-22(h) All 2'-deoxynucleosides l(g)-10(g)

341 l(i)-22(i) All 2'-deoxynucleosides l(g)-10(g)

342 l(a)-22(a) All 2'-deoxynucleosides 1(h)- 10(h)

343 l(b)-22(b) All 2'-deoxynucleosides 1(h)- 10(h)

344 l(c)-22(c) All 2'-deoxynucleosides 1(h)- 10(h)

345 l(d)-22(d) All 2'-deoxynucleosides 1(h)- 10(h)

346 l(e)-22(e) All 2'-deoxynucleosides 1(h)- 10(h)

347 l(f)-22(f) All 2'-deoxynucleosides 1(h)- 10(h)

348 l(g)-22(g) All 2'-deoxynucleosides 1(h)- 10(h)

349 l(h)-22(h) All 2'-deoxynucleosides 1(h)- 10(h)

350 l(i)-22(i) All 2'-deoxynucleosides 1(h)- 10(h)

351 l(a)-22(a) All 2'-deoxynucleosides l(i)- 10(i)

352 l(b)-22(b) All 2'-deoxynucleosides l(i)- 10(i)

353 l(c)-22(c) All 2'-deoxynucleosides l(i)- 10(i)

354 l(d)-22(d) All 2'-deoxynucleosides l(i)- 10(i)

355 l(e)-22(e) All 2'-deoxynucleosides l(i)- 10(i)

356 l(f)-22(f) All 2'-deoxynucleosides l(i)- 10(i)

357 l(g)-22(g) All 2'-deoxynucleosides l(i)- 10(i)

358 l(h)-22(h) All 2'-deoxynucleosides l(i)- 10(i)

359 l(i)-22(i) All 2'-deoxynucleosides l(i)- 10(i)

360 l(a-l) All 2'-deoxynucleosides l(a-l)

361 2(a-l) All 2'-deoxynucleosides l(a-l)

362 3(a-l) All 2'-deoxynucleosides l(a-l)

363 4(a-l) All 2'-deoxynucleosides l(a-l)

364 5(a-l) All 2'-deoxynucleosides l(a-l)

365 6(a-l) All 2'-deoxynucleosides l(a-l)

366 7(a-l) All 2'-deoxynucleosides l(a-l)

367 8(a-l) All 2'-deoxynucleosides l(a-l) 368 9(a-l) All 2'-deoxynucleosides l(a-l)

369 10(a-l) All 2'-deoxynucleosides l(a-l)

370 l l(a-l) All 2'-deoxynucleosides l(a-l)

371 12(a-l) All 2'-deoxynucleosides l(a-l)

372 13(a-l) All 2'-deoxynucleosides l(a-l)

373 14(a-l) All 2'-deoxynucleosides l(a-l)

374 15(a-l) All 2'-deoxynucleosides l(a-l)

375 16(a-l) All 2'-deoxynucleosides l(a-l)

376 17(a-l) All 2'-deoxynucleosides l(a-l)

377 18(a-l) All 2'-deoxynucleosides l(a-l)

378 19(a-l) All 2'-deoxynucleosides l(a-l)

379 20(a-l) All 2'-deoxynucleosides l(a-l)

380 21(a-l) All 2'-deoxynucleosides l(a-l)

381 22(a-l) All 2'-deoxynucleosides l(a-l)

382 l(a-l) All 2'-deoxynucleosides 2(a-l)

383 2(a-l) All 2'-deoxynucleosides 2(a-l)

384 3(a-l) All 2'-deoxynucleosides 2(a-l)

385 4(a-l) All 2'-deoxynucleosides 2(a-l)

386 5(a-l) All 2'-deoxynucleosides 2(a-l)

387 6(a-l) All 2'-deoxynucleosides 2(a-l)

388 7(a-l) All 2'-deoxynucleosides 2(a-l)

389 8(a-l) All 2'-deoxynucleosides 2(a-l)

390 9(a-l) All 2'-deoxynucleosides 2(a-l)

391 10(a-l) All 2'-deoxynucleosides 2(a-l)

392 l l(a-l) All 2'-deoxynucleosides 2(a-l)

393 12(a-l) All 2'-deoxynucleosides 2(a-l)

394 13(a-l) All 2'-deoxynucleosides 2(a-l)

395 14(a-l) All 2'-deoxynucleosides 2(a-l)

396 15(a-l) All 2'-deoxynucleosides 2(a-l)

397 16(a-l) All 2'-deoxynucleosides 2(a-l)

398 17(a-l) All 2'-deoxynucleosides 2(a-l)

399 18(a-l) All 2'-deoxynucleosides 2(a-l)

400 19(a-l) All 2'-deoxynucleosides 2(a-l)

401 20(a-l) All 2'-deoxynucleosides 2(a-l)

402 21(a-l) All 2'-deoxynucleosides 2(a-l)

403 22(a-l) All 2'-deoxynucleosides 2(a-l)

404 l(a-l) All 2'-deoxynucleosides 3(a-l)

405 2(a-l) All 2'-deoxynucleosides 3(a-l)

406 3(a-l) All 2'-deoxynucleosides 3(a-l)

407 4(a-l) All 2'-deoxynucleosides 3(a-l)

408 5(a-l) All 2'-deoxynucleosides 3(a-l)

409 6(a-l) All 2'-deoxynucleosides 3(a-l)

410 7(a-l) All 2'-deoxynucleosides 3(a-l)

41 1 8(a-l) All 2'-deoxynucleosides 3(a-l)

412 9(a-l) All 2'-deoxynucleosides 3(a-l) 413 10(a-l) All 2'-deoxynucleosides 3(a-l)

414 l l(a-l) All 2'-deoxynucleosides 3(a-l)

415 12(a-l) All 2'-deoxynucleosides 3(a-l)

416 13(a-l) All 2'-deoxynucleosides 3(a-l)

417 14(a-l) All 2'-deoxynucleosides 3(a-l)

418 15(a-l) All 2'-deoxynucleosides 3(a-l)

419 16(a-l) All 2'-deoxynucleosides 3(a-l)

420 17(a-l) All 2'-deoxynucleosides 3(a-l)

421 18(a-l) All 2'-deoxynucleosides 3(a-l)

422 19(a-l) All 2'-deoxynucleosides 3(a-l)

423 20(a-l) All 2'-deoxynucleosides 3(a-l)

424 21(a-l) All 2'-deoxynucleosides 3(a-l)

425 22(a-l) All 2'-deoxynucleosides 3(a-l)

426 l(a-l) All 2'-deoxynucleosides 4(a-l)

427 2(a-l) All 2'-deoxynucleosides 4(a-l)

428 3(a-l) All 2'-deoxynucleosides 4(a-l)

429 4(a-l) All 2'-deoxynucleosides 4(a-l)

430 5(a-l) All 2'-deoxynucleosides 4(a-l)

431 6(a-l) All 2'-deoxynucleosides 4(a-l)

432 7(a-l) All 2'-deoxynucleosides 4(a-l)

433 8(a-l) All 2'-deoxynucleosides 4(a-l)

434 9(a-l) All 2'-deoxynucleosides 4(a-l)

435 10(a-l) All 2'-deoxynucleosides 4(a-l)

436 l l(a-l) All 2'-deoxynucleosides 4(a-l)

437 12(a-l) All 2'-deoxynucleosides 4(a-l)

438 13(a-l) All 2'-deoxynucleosides 4(a-l)

439 14(a-l) All 2'-deoxynucleosides 4(a-l)

440 15(a-l) All 2'-deoxynucleosides 4(a-l)

441 16(a-l) All 2'-deoxynucleosides 4(a-l)

442 17(a-l) All 2'-deoxynucleosides 4(a-l)

443 18(a-l) All 2'-deoxynucleosides 4(a-l)

444 19(a-l) All 2'-deoxynucleosides 4(a-l)

445 20(a-l) All 2'-deoxynucleosides 4(a-l)

446 21(a-l) All 2'-deoxynucleosides 4(a-l)

447 22(a-l) All 2'-deoxynucleosides 4(a-l)

448 l(a-l) All 2'-deoxynucleosides 5(a-l)

449 2(a-l) All 2'-deoxynucleosides 5(a-l)

450 3(a-l) All 2'-deoxynucleosides 5(a-l)

451 4(a-l) All 2'-deoxynucleosides 5(a-l)

452 5(a-l) All 2'-deoxynucleosides 5(a-l)

453 6(a-l) All 2'-deoxynucleosides 5(a-l)

454 7(a-l) All 2'-deoxynucleosides 5(a-l)

455 8(a-l) All 2'-deoxynucleosides 5(a-l)

456 9(a-l) All 2'-deoxynucleosides 5(a-l)

457 10(a-l) All 2'-deoxynucleosides 5(a-l) 458 l l(a-l) All 2'-deoxynucleosides 5(a-l)

459 12(a-l) All 2'-deoxynucleosides 5(a-l)

460 13(a-l) All 2'-deoxynucleosides 5(a-l)

461 14(a-l) All 2'-deoxynucleosides 5(a-l)

462 15(a-l) All 2'-deoxynucleosides 5(a-l)

463 16(a-l) All 2'-deoxynucleosides 5(a-l)

464 17(a-l) All 2'-deoxynucleosides 5(a-l)

465 18(a-l) All 2'-deoxynucleosides 5(a-l)

466 19(a-l) All 2'-deoxynucleosides 5(a-l)

467 20(a-l) All 2'-deoxynucleosides 5(a-l)

468 21(a-l) All 2'-deoxynucleosides 5(a-l)

469 22(a-l) All 2'-deoxynucleosides 5(a-l)

470 l(a-l) All 2'-deoxynucleosides 6(a-l)

471 2(a-l) All 2'-deoxynucleosides 6(a-l)

472 3(a-l) All 2'-deoxynucleosides 6(a-l)

473 4(a-l) All 2'-deoxynucleosides 6(a-l)

474 5(a-l) All 2'-deoxynucleosides 6(a-l)

475 6(a-l) All 2'-deoxynucleosides 6(a-l)

476 7(a-l) All 2'-deoxynucleosides 6(a-l)

477 8(a-l) All 2'-deoxynucleosides 6(a-l)

478 9(a-l) All 2'-deoxynucleosides 6(a-l)

479 10(a-l) All 2'-deoxynucleosides 6(a-l)

480 l l(a-l) All 2'-deoxynucleosides 6(a-l)

481 12(a-l) All 2'-deoxynucleosides 6(a-l)

482 13(a-l) All 2'-deoxynucleosides 6(a-l)

483 14(a-l) All 2'-deoxynucleosides 6(a-l)

484 15(a-l) All 2'-deoxynucleosides 6(a-l)

485 16(a-l) All 2'-deoxynucleosides 6(a-l)

486 17(a-l) All 2'-deoxynucleosides 6(a-l)

487 18(a-l) All 2'-deoxynucleosides 6(a-l)

488 19(a-l) All 2'-deoxynucleosides 6(a-l)

489 20(a-l) All 2'-deoxynucleosides 6(a-l)

490 21(a-l) All 2'-deoxynucleosides 6(a-l)

491 22(a-l) All 2'-deoxynucleosides 6(a-l)

492 l(a-l) All 2'-deoxynucleosides 7(a-l)

493 2(a-l) All 2'-deoxynucleosides 7(a-l)

494 3(a-l) All 2'-deoxynucleosides 7(a-l)

495 4(a-l) All 2'-deoxynucleosides 7(a-l)

496 5(a-l) All 2'-deoxynucleosides 7(a-l)

497 6(a-l) All 2'-deoxynucleosides 7(a-l)

498 7(a-l) All 2'-deoxynucleosides 7(a-l)

499 8(a-l) All 2'-deoxynucleosides 7(a-l)

500 9(a-l) All 2'-deoxynucleosides 7(a-l)

501 10(a-l) All 2'-deoxynucleosides 7(a-l)

502 l l(a-l) All 2'-deoxynucleosides 7(a-l) 503 12(a-l) All 2'-deoxynucleosides 7(a-l)

504 13(a-l) All 2'-deoxynucleosides 7(a-l)

505 14(a-l) All 2'-deoxynucleosides 7(a-l)

506 15(a-l) All 2'-deoxynucleosides 7(a-l)

507 16(a-l) All 2'-deoxynucleosides 7(a-l)

508 17(a-l) All 2'-deoxynucleosides 7(a-l)

509 18(a-l) All 2'-deoxynucleosides 7(a-l)

510 19(a-l) All 2'-deoxynucleosides 7(a-l)

51 1 20(a-l) All 2'-deoxynucleosides 7(a-l)

512 21(a-l) All 2'-deoxynucleosides 7(a-l)

513 22(a-l) All 2'-deoxynucleosides 7(a-l)

514 l(a-l) All 2'-deoxynucleosides 8(a-l)

515 2(a-l) All 2'-deoxynucleosides 8(a-l)

516 3(a-l) All 2'-deoxynucleosides 8(a-l)

517 4(a-l) All 2'-deoxynucleosides 8(a-l)

518 5(a-l) All 2'-deoxynucleosides 8(a-l)

519 6(a-l) All 2'-deoxynucleosides 8(a-l)

520 7(a-l) All 2'-deoxynucleosides 8(a-l)

521 8(a-l) All 2'-deoxynucleosides 8(a-l)

522 9(a-l) All 2'-deoxynucleosides 8(a-l)

523 10(a-l) All 2'-deoxynucleosides 8(a-l)

524 l l(a-l) All 2'-deoxynucleosides 8(a-l)

525 12(a-l) All 2'-deoxynucleosides 8(a-l)

526 13(a-l) All 2'-deoxynucleosides 8(a-l)

527 14(a-l) All 2'-deoxynucleosides 8(a-l)

528 15(a-l) All 2'-deoxynucleosides 8(a-l)

529 16(a-l) All 2'-deoxynucleosides 8(a-l)

530 17(a-l) All 2'-deoxynucleosides 8(a-l)

531 18(a-l) All 2'-deoxynucleosides 8(a-l)

532 19(a-l) All 2'-deoxynucleosides 8(a-l)

533 20(a-l) All 2'-deoxynucleosides 8(a-l)

534 21(a-l) All 2'-deoxynucleosides 8(a-l)

535 22(a-l) All 2'-deoxynucleosides 8(a-l)

536 l(a-l) All 2'-deoxynucleosides 9(a-l)

537 2(a-l) All 2'-deoxynucleosides 9(a-l)

538 3(a-l) All 2'-deoxynucleosides 9(a-l)

539 4(a-l) All 2'-deoxynucleosides 9(a-l)

540 5(a-l) All 2'-deoxynucleosides 9(a-l)

541 6(a-l) All 2'-deoxynucleosides 9(a-l)

542 7(a-l) All 2'-deoxynucleosides 9(a-l)

543 8(a-l) All 2'-deoxynucleosides 9(a-l)

544 9(a-l) All 2'-deoxynucleosides 9(a-l)

545 10(a-l) All 2'-deoxynucleosides 9(a-l)

546 l l(a-l) All 2'-deoxynucleosides 9(a-l)

547 12(a-l) All 2'-deoxynucleosides 9(a-l) 548 13(a-l) All 2'-deoxynucleosides 9(a-l)

549 14(a-l) All 2'-deoxynucleosides 9(a-l)

550 15(a-l) All 2'-deoxynucleosides 9(a-l)

551 16(a-l) All 2'-deoxynucleosides 9(a-l)

552 17(a-l) All 2'-deoxynucleosides 9(a-l)

553 18(a-l) All 2'-deoxynucleosides 9(a-l)

554 19(a-l) All 2'-deoxynucleosides 9(a-l)

555 20(a-l) All 2'-deoxynucleosides 9(a-l)

556 21(a-l) All 2'-deoxynucleosides 9(a-l)

557 22(a-l) All 2'-deoxynucleosides 9(a-l)

558 l(a-l) All 2'-deoxynucleosides 10(a-l)

559 2(a-l) All 2'-deoxynucleosides 10(a-l)

560 3(a-l) All 2'-deoxynucleosides 10(a-l)

561 4(a-l) All 2'-deoxynucleosides 10(a-l)

562 5(a-l) All 2'-deoxynucleosides 10(a-l)

563 6(a-l) All 2'-deoxynucleosides 10(a-l)

564 7(a-l) All 2'-deoxynucleosides 10(a-l)

565 8(a-l) All 2'-deoxynucleosides 10(a-l)

566 9(a-l) All 2'-deoxynucleosides 10(a-l)

567 10(a-l) All 2'-deoxynucleosides 10(a-l)

568 l l(a-l) All 2'-deoxynucleosides 10(a-l)

569 12(a-l) All 2'-deoxynucleosides 10(a-l)

570 13(a-l) All 2'-deoxynucleosides 10(a-l)

571 14(a-l) All 2'-deoxynucleosides 10(a-l)

572 15(a-l) All 2'-deoxynucleosides 10(a-l)

573 16(a-l) All 2'-deoxynucleosides 10(a-l)

574 17(a-l) All 2'-deoxynucleosides 10(a-l)

575 18(a-l) All 2'-deoxynucleosides 10(a-l)

576 19(a-l) All 2'-deoxynucleosides 10(a-l)

577 20(a-l) All 2'-deoxynucleosides 10(a-l)

578 21(a-l) All 2'-deoxynucleosides 10(a-l)

579 22(a-l) All 2'-deoxynucleosides 10(a-l)

580 l(j)-22(j) All 2'-deoxynucleosides l(a)-10(a)

581 l(k)-22(k) All 2'-deoxynucleosides l(a)-10(a)

582 1(1)-22(1) All 2'-deoxynucleosides l(a)-10(a)

583 l(j)-22(j) All 2'-deoxynucleosides l(b)-10(b)

584 l(k)-22(k) All 2'-deoxynucleosides l(b)-10(b)

585 1(1)-22(1) All 2'-deoxynucleosides l(b)-10(b)

586 l(j)-22(j) All 2'-deoxynucleosides l(c)-10(c)

587 l(k)-22(k) All 2'-deoxynucleosides l(c)-10(c)

588 1(1)-22(1) All 2'-deoxynucleosides l(c)-10(c)

589 l(j)-22(j) All 2'-deoxynucleosides l(d)-10(d)

590 l(k)-22(k) All 2'-deoxynucleosides l(d)-10(d)

591 1(1)-22(1) All 2'-deoxynucleosides l(d)-10(d)

592 l(j)-22(j) All 2'-deoxynucleosides l(e)-10(e) 593 l(k)-22(k) All 2'-deoxynucleosides l(e)-10(e)

594 1(1)-22(1) All 2'-deoxynucleosides l(e)-10(e)

595 l(j)-22(j) All 2'-deoxynucleosides l(f)-10(f)

596 l(k)-22(k) All 2'-deoxynucleosides l(f)-10(f)

597 1(1)-22(1) All 2'-deoxynucleosides l(f)-10(f)

598 l(j)-22(j) All 2'-deoxynucleosides l(g)-10(g)

599 l(k)-22(k) All 2'-deoxynucleosides l(g)-10(g)

600 1(1)-22(1) All 2'-deoxynucleosides l(g)-10(g)

601 l(j)-22(j) All 2'-deoxynucleosides l(h)-10(h)

602 l(k)-22(k) All 2'-deoxynucleosides l(h)-10(h)

603 1(1)-22(1) All 2'-deoxynucleosides l(h)-10(h)

604 l(j)-22(j) All 2'-deoxynucleosides l(i)- 10(i)

605 l(k)-22(k) All 2'-deoxynucleosides l(i)- 10(i)

606 1(1)-22(1) All 2'-deoxynucleosides l(i)- 10(i)

607 l(j)-22(j) All 2'-deoxynucleosides i(j)-ioa)

608 l(k)-22(k) All 2'-deoxynucleosides i(j)-ioa)

609 1(1)-22(1) All 2'-deoxynucleosides i(j)-ioa)

610 l(j)-22(j) All 2'-deoxynucleosides l(k)-10(k)

61 1 l(k)-22(k) All 2'-deoxynucleosides l(k)-10(k)

612 1(1)-22(1) All 2'-deoxynucleosides l(k)-10(k)

612 l(j)-22(j) All 2'-deoxynucleosides 1(1)- 10(1)

614 l(k)-22(k) All 2'-deoxynucleosides 1(1)- 10(1)

615 1(1)-22(1) All 2'-deoxynucleosides 1(1)- 10(1)

616 lk All 2'-deoxynucleosides lm

In certain embodiments, a gapmer comprises a 5 '-wing selected from among the 5 '-wings provided herein and any 3 '-wing. In certain embodiments, a gapmer comprises a 5 '-wing selected from among l(a-i) to 22(a-i). In certain embodiments, a gapmer comprises a 5'-wing selected from among l(a-l) to 22(a-l). In certain embodiments, a gapmer comprises a 3 '-wing selected from among the 3 '-wings provided herein and any 5'-wing. In certain embodiments, a gapmer comprises a 3'-wing selected from among l(a-i) to 10(a-i). In certain embodiments, a gapmer comprises a 3 '-wing selected from among l(a-l) to 10(a-l).

In certain embodiments, a gapmer has a sugar motif other than: E-K-K-(D)9-K-K-E; E-E-E-E-K-(D)9- E-E-E-E-E; E-K-K-K-(D)9-K-K-K-E; K-E-E-K-(D)9-K-E-E-K; K-D-D-K-(D)9-K-D-D-K; K-E-K-E-K-(D)9- K-E-K-E-K; K-D-K-D-K-(D)9-K-D-K-D-K; E-K-E-K-(D)9-K-E-K-E; E-E-E-E-E-K-(D)8-E-E-E-E-E; or E- K-E-K-E-(D)9-E-K-E-K-E. In certain embodiments, a gapmer not having one of the above motifs has a sugar motif of Formula I. In certain embodiments, a gapmer not having one of the above motifs has a sugar motif selected from motifs 1-58. In certain embodiments, a gapmer not having one of the above motifs has a sugar motif of Formula I and selected from sugar motifs 1-58. In certain embodiments, a gapmer not having one of the above motifs has a sugar motif of Formula II. In certain embodiments, a gapmer not having one of the above motifs has a sugar motif selected from motifs 1-615. In certain embodiments, a gapmer not having one of the above motifs has a sugar motif of Formula II and selected from sugar motifs 1-615. In certain embodiments a gapmer comprises a A-(D)4-A-(D)4-A-(D)4-AA motif. In certain embodiments a gapmer comprises a B-(D)4-A-(D)4-A-(D)4-AA motif. In certain embodiments a gapmer comprises a A-(D)4-B-(D)4-A-(D)4-AA motif. In certain embodiments a gapmer comprises a A-(D)4-A-(D)4- B-(D)4-AA motif. In certain embodiments a gapmer comprises a A-(D)4-A-(D)4-A-(D)4-BA motif. In certain embodiments a gapmer comprises a A-(D)4-A-(D)4-A-(D)4-BB motif. In certain embodiments a gapmer comprises a K-(D)4-K-(D)4-K-(D)4-K-E motif.

Certain mtemucleoside Linkage Motifs

In certain embodiments, oligonucleotides comprise modified mtemucleoside linkages arranged along the oligonucleotide or region thereof in a defined pattern or modified mtemucleoside linkage motif. In certain embodiments, mtemucleoside linkages are arranged in a gapped motif, as described above for sugar modification motif. In such embodiments, the mtemucleoside linkages in each of two wing regions are different from the mtemucleoside linkages in the gap region. In certain embodiments the mtemucleoside linkages in the wings are phosphodiester and the mtemucleoside linkages in the gap are phosphorothioate. The sugar modification motif is independently selected, so such oligonucleotides having a gapped mtemucleoside linkage motif may or may not have a gapped sugar modification motif and if it does have a gapped sugar motif, the wing and gap lengths may or may not be the same.

In certain embodiments, oligonucleotides comprise a region having an alternating mtemucleoside linkage motif. In certain embodiments, oligonucleotides of the present invention comprise a region of uniformly modified mtemucleoside linkages. In certain such embodiments, the oligonucleotide comprises a region that is uniformly linked by phosphorothioate mtemucleoside linkages. In certain embodiments, the oligonucleotide is uniformly linked by phosphorothioate. In certain embodiments, each mtemucleoside linkage of the oligonucleotide is selected from phosphodiester and phosphorothioate. In certain

embodiments, each mtemucleoside linkage of the oligonucleotide is selected from phosphodiester and phosphorothioate and at least one mtemucleoside linkage is phosphorothioate.

In certain embodiments, the oligonucleotide comprises at least 6 phosphorothioate mtemucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 8 phosphorothioate mtemucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 10 phosphorothioate mtemucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 6 consecutive phosphorothioate mtemucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 8 consecutive phosphorothioate mtemucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 10 consecutive phosphorothioate mtemucleoside linkages. In certain embodiments, the oligonucleotide comprises at least block of at least one 12 consecutive phosphorothioate mtemucleoside linkages. In certain such embodiments, at least one such block is located at the 3 ' end of the oligonucleotide. In certain such embodiments, at least one such block is located within 3 nucleosides of the 3 ' end of the oligonucleotide. Certain Nucleobase Modification Motifs

In certain embodiments, oligonucleotides comprise chemical modifications to nucleobases arranged along the oligonucleotide or region thereof in a defined pattern or nucleobases modification motif. In certain such embodiments, nucleobase modifications are arranged in a gapped motif. In certain embodiments, nucleobase modifications are arranged in an alternating motif. In certain embodiments, each nucleobase is modified. In certain embodiments, none of the nucleobases is chemically modified.

In certain embodiments, oligonucleotides comprise a block of modified nucleobases. In certain such embodiments, the block is at the 3 '-end of the oligonucleotide. In certain embodiments the block is within 3 nucleotides of the 3 '-end of the oligonucleotide. In certain such embodiments, the block is at the 5 '-end of the oligonucleotide. In certain embodiments the block is within 3 nucleotides of the 5 '-end of the oligonucleotide.

In certain embodiments, nucleobase modifications are a function of the natural base at a particular position of an oligonucleotide. For example, in certain embodiments each purine or each pyrimidine in an oligonucleotide is modified. In certain embodiments, each adenine is modified. In certain embodiments, each guanine is modified. In certain embodiments, each thymine is modified. In certain embodiments, each cytosine is modified. In certain embodiments, each uracil is modified.

In certain embodiments, some, all, or none of the cytosine moieties in an oligonucleotide are 5- methyl cytosine moieties. Herein, 5-methyl cytosine is not a "modified nucleobase." Accordingly, unless otherwise indicated, unmodified nucleobases include both cytosine residues having a 5-methyl and those lacking a 5 methyl. In certain embodiments, the methylation state of all or some cytosine nucleobases is specified.

Certain Overall Lengths

In certain embodiments, the present invention provides oligomeric compounds including

oligonucleotides of any of a variety of ranges of lengths. In certain embodiments, the invention provides oligomeric compounds or oligonucleotides consisting of X to Y linked nucleosides, where X represents the fewest number of nucleosides in the range and Y represents the largest number of nucleosides in the range. In certain such embodiments, X and Y are each independently selected from 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50; provided that X<Y. For example, in certain embodiments, the invention provides oligomeric compounds which comprise oligonucleotides consisting of 8 to 9, 8 to 10, 8 to 1 1, 8 to 12, 8 to 13, 8 to 14, 8 to 15, 8 to 16, 8 to 17, 8 to 18, 8 to 19, 8 to 20, 8 to 21, 8 to 22, 8 to 23, 8 to 24, 8 to 25, 8 to 26, 8 to 27, 8 to 28, 8 to 29, 8 to 30, 9 to 10, 9 to 1 1, 9 to 12, 9 to 13, 9 to 14, 9 to 15, 9 to 16, 9 to 17, 9 to 18, 9 to 19, 9 to 20, 9 to 21, 9 to 22, 9 to 23, 9 to 24, 9 to 25, 9 to 26, 9 to 27, 9 to 28, 9 to 29, 9 to 30, 10 to 1 1, 10 to 12, 10 to 13, 10 to 14, 10 to 15, 10 to 16, 10 to 17, 10 to 18, 10 to 19, 10 to 20, 10 to 21, 10 to 22, 10 to 23, 10 to 24, 10 to 25, 10 to 26, 10 to 27, 10 to 28, 10 to 29, 10 to 30, 1 1 to 12, 11 to 13, 1 1 to 14, 1 1 to 15, 1 1 to 16, 1 1 to 17, 1 1 to 18, 1 1 to 19, 1 1 to 20, 1 1 to 21, 1 1 to 22, 1 1 to 23, 1 1 to 24, 1 1 to 25, 1 1 to 26, 1 1 to 27,

1 1 to 28, 11 to 29, 1 1 to 30, 12 to 13, 12 to 14, 12 to 15, 12 to 16, 12 to 17, 12 to 18, 12 to 19, 12 to 20, 12 to 21, 12 to 22, 12 to 23, 12 to 24, 12 to 25, 12 to 26, 12 to 27, 12 to 28, 12 to 29, 12 to 30, 13 to 14, 13 to 15,

13 to 16, 13 to 17, 13 to 18, 13 to 19, 13 to 20, 13 to 21, 13 to 22, 13 to 23, 13 to 24, 13 to 25, 13 to 26, 13 to

27, 13 to 28, 13 to 29, 13 to 30, 14 to 15, 14 to 16, 14 to 17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22,

14 to 23, 14 to 24, 14 to 25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, 15 to 16, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to 23, 15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to 30,

16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16 to 23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to

28, 16 to 29, 16 to 30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to 24, 17 to 25, 17 to 26,

17 to 27, 17 to 28, 17 to 29, 17 to 30, 18 to 19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24, 18 to 25, 18 to

26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to 20, 19 to 21, 19 to 22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19 to 29, 19 to 28, 19 to 29, 19 to 30, 20 to 21, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 20 to 26, 20 to 27, 20 to 28, 20 to 29, 20 to 30, 21 to 22, 21 to 23, 21 to 24, 21 to 25, 21 to 26, 21 to 27, 21 to 28, 21 to 29, 21 to 30, 22 to 23, 22 to 24, 22 to 25, 22 to 26, 22 to 27, 22 to 28, 22 to 29, 22 to 30, 23 to 24, 23 to 25, 23 to 26, 23 to

27, 23 to 28, 23 to 29, 23 to 30, 24 to 25, 24 to 26, 24 to 27, 24 to 28, 24 to 29, 24 to 30, 25 to 26, 25 to 27, 25 to 28, 25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to 29, 26 to 30, 27 to 28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to 30 linked nucleosides. In embodiments where the number of nucleosides of an oligomeric compound or oligonucleotide is limited, whether to a range or to a specific number, the oligomeric compound or oligonucleotide may, nonetheless further comprise additional other substituents. For example, an oligonucleotide comprising 8-30 nucleosides excludes oligonucleotides having 31 nucleosides, but, unless otherwise indicated, such an oligonucleotide may further comprise, for example one or more conjugates, terminal groups, or other substituents. In certain embodiments, a gapmer oligonucleotide has any of the above lengths.

In certain embodiments, any of the gapmer motifs provided above, including but not limited to gapmer motifs 1-278 provided in Tables 3 and 4, may have any of the above lengths. One of skill in the art will appreciate that certain lengths may not be possible for certain motifs. For example: a gapmer having a 5 '-wing region consisting of four nucleotides, a gap consisting of at least six nucleotides, and a 3 '-wing region consisting of three nucleotides cannot have an overall length less than 13 nucleotides. Thus, one would understand that the lower length limit is 13 and that the limit of 10 in "10-20" has no effect in that embodiment.

Further, where an oligonucleotide is described by an overall length range and by regions having specified lengths, and where the sum of specified lengths of the regions is less than the upper limit of the overall length range, the oligonucleotide may have additional nucleosides, beyond those of the specified regions, provided that the total number of nucleosides does not exceed the upper limit of the overall length range. For example, an oligonucleotide consisting of 20-25 linked nucleosides comprising a 5 '-wing consisting of 5 linked nucleosides; a 3 '-wing consisting of 5 linked nucleosides and a central gap consisting of 10 linked nucleosides (5+5+10=20) may have up to 5 nucleosides that are not part of the 5 '-wing, the 3'- wing, or the gap (before reaching the overall length limitation of 25). Such additional nucleosides may be 5' of the 5 '-wing and/or 3' of the 3' wing.

Certain Oligonucleotides

In certain embodiments, oligonucleotides of the present invention are characterized by their sugar motif, internucleoside linkage motif, nucleobase modification motif and overall length. In certain embodiments, such parameters are each independent of one another. Thus, each internucleoside linkage of an oligonucleotide having a gapmer sugar motif may be modified or unmodified and may or may not follow the gapmer modification pattern of the sugar modifications. Thus, the internucleoside linkages within the wing regions of a sugar-gapmer may be the same or different from one another and may be the same or different from the internucleoside linkages of the gap region. Likewise, such sugar-gapmer oligonucleotides may comprise one or more modified nucleobase independent of the gapmer pattern of the sugar modifications. One of skill in the art will appreciate that such motifs may be combined to create a variety of

oligonucleotides, such as those provided in the non-limiting Table 5 below.

Table 10

Certain Oligonucleotides

Figure imgf000102_0001
Gapmer of Formula I and having uniform PS uniform unmodified motif selected from 1-58

Gapmer motif selected from 1 - uniform PS uniform unmodified 278

Gapmer of Formula I uniform PS uniform unmodified

Gapmer motif selected from 1 - uniform PS uniform unmodified 359

Gapmer motif selected from 1 - 2-14-2 gapmer: PO in uniform unmodified 359 wings and PS in gap

Gapmer motif selected from 1 - uniform PS uniform unmodified; 359 all C's are 5-meC

Gapmer of Formula II uniform PS uniform unmodified;

no Cs are 5-meC)

Gapmer of Formula II uniform PS uniform unmodified;

at least one nucleobase is a 5-meC

Gapmer of Formula II and having uniform PS uniform unmodified motif selected from 1-359

Gapmer of Formula II and having uniform PO uniform unmodified motif selected from 1-359

Gapmer motif selected from 1 - uniform PS uniform unmodified 359

Gapmer of Formula II uniform PS uniform unmodified

Gapmer of Formula I and having uniform PS uniform unmodified motif selected from 1-359

Gapmer motif selected from 1 - uniform PS uniform unmodified 359

Gapmer of Formula II uniform PS uniform unmodified

Gapmer motif selected from 1 - uniform PS uniform unmodified 615

Gapmer motif selected from 1 - 2-14-2 gapmer: PO in uniform unmodified 615 wings and PS in gap

Gapmer motif selected from 1 - uniform PS uniform unmodified; 615 all C's are 5-meC

Gapmer of Formula I and having uniform PS uniform unmodified motif selected from 1-615

Gapmer of Formula I and having uniform PO uniform unmodified motif selected from 1-615

Gapmer motif selected from 1 - uniform PS uniform unmodified 615

Gapmer of Formula I and having uniform PS uniform unmodified motif selected from 1-615

Gapmer motif selected from 1 - uniform PS uniform unmodified 615 The above table is intended only to illustrate and not to limit the various combinations of the parameters of oligonucleotides of the present invention. Herein if a description of an oligonucleotide or oligomeric compound is silent with respect to one or more parameter, such parameter is not limited. Thus, an oligomeric compound described only as having a gapmer sugar motif without further description may have any length, internucleoside linkage motif, and nucleobase modification motif. Unless otherwise indicated, all chemical modifications are independent of nucleobase sequence.

Certain Conjugate Groups

In certain embodiments, oligomeric compounds are modified by attachment of one or more conjugate groups. In general, conjugate groups modify one or more properties of the attached oligomeric compound including but not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, cellular distribution, cellular uptake, charge and clearance. Conjugate groups are routinely used in the chemical arts and are linked directly or via an optional conjugate linking moiety or conjugate linking group to a parent compound such as an oligomeric compound, such as an oligonucleotide. Conjugate groups includes without limitation, intercalators, reporter molecules, polyamines, polyamides, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipids, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins and dyes. Certain conjugate groups have been described previously, for example: cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Let., 1994, 4, 1053- 1060), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Let., 1993, 3, 2765-2770), a thiocholesterol

(Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, e.g., do-decan-diol or undecyl residues (Saison-Behmoaras et al., EMBO J., 1991 , 10, 1 1 1 1- 1 1 18; Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al., Biochimie, 1993, 75, 49-54), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethyl-ammonium l ,2-di-0-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651 -3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651 -3654), a palmityl moiety (Mishra et al., Biochim.

Biophys. Acta, 1995, 1264, 229-237), or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol. Exp. Ther., 1996, 277, 923-937).

In certain embodiments, a conjugate group comprises an active drug substance, for example, aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansylsarcosine, 2,3,5-triiodobenzoic acid, flufenamic acid, folinic acid, a benzothiadiazide, chlorothiazide, a diazepine, indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, an antidiabetic, an antibacterial or an antibiotic.

In certain embodiments, conjugate groups are directly attached to oligonucleotides in oligomeric compounds. In certain embodiments, conjugate groups are attached to oligonucleotides by a conjugate linking group. In certain such embodiments, conjugate linking groups, including, but not limited to, bifunctional linking moieties such as those known in the art are amenable to the compounds provided herein. Conjugate linking groups are useful for attachment of conjugate groups, such as chemical stabilizing groups, functional groups, reporter groups and other groups to selective sites in a parent compound such as for example an oligomeric compound. In general a bifunctional linking moiety comprises a hydrocarbyl moiety having two functional groups. One of the functional groups is selected to bind to a parent molecule or compound of interest and the other is selected to bind essentially any selected group such as chemical functional group or a conjugate group. In some embodiments, the conjugate linker comprises a chain structure or an oligomer of repeating units such as ethylene glycol or amino acid units. Examples of functional groups that are routinely used in a bifunctional linking moiety include, but are not limited to, electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In some embodiments, bifunctional linking moieties include amino, hydroxyl, carboxylic acid, thiol, unsaturations (e.g., double or triple bonds), and the like.

Some nonlimiting examples of conjugate linking moieties include pyrrolidine, 8-amino-3,6- dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC) and 6- aminohexanoic acid (AHEX or AHA). Other linking groups include, but are not limited to, substituted Cp Cio alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted C2-C10 alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.

Conjugate groups may be attached to either or both ends of an oligonucleotide (terminal conjugate groups) and/or at any internal position.

In certain embodiments, conjugate groups are at the 3 '-end of an oligonucleotide of an oligomeric compound. In certain embodiments, conjugate groups are near the 3 '-end. In certain embodiments, conjugates are attached at the 3 'end of an oligomeric compound, but before one or more terminal group nucleosides. In certain embodiments, conjugate groups are placed within a terminal group.

In certain embodiments, the present invention provides oligomeric compounds. In certain embodiments, oligomeric compounds comprise an oligonucleotide. In certain embodiments, an oligomeric compound comprises an oligonucleotide and one or more conjugate and/or terminal groups. Such conjugate and/or terminal groups may be added to oligonucleotides having any of the chemical motifs discussed above. Thus, for example, an oligomeric compound comprising an oligonucleotide having region of alternating nucleosides may comprise a terminal group.

Antisense Compounds

In certain embodiments, oligomeric compounds of the present invention are antisense compounds. Such antisense compounds are capable of hybridizing to a target nucleic acid, resulting in at least one antisense activity. In certain embodiments, antisense compounds specifically hybridize to one or more target nucleic acid. In certain embodiments, a specifically hybridizing antisense compound has a nucleobase sequence comprising a region having sufficient complementarity to a target nucleic acid to allow

hybridization and result in antisense activity and insufficient complementarity to any non-target so as to avoid non-specific hybridization to any non-target nucleic acid sequences under conditions in which specific hybridization is desired (e.g., under physiological conditions for in vivo or therapeutic uses, and under conditions in which assays are performed in the case of in vitro assays).

In certain embodiments, the present invention provides antisense compounds comprising

oligonucleotides that are fully complementary to the target nucleic acid over the entire length of the oligonucleotide. In certain embodiments, oligonucleotides are 99% complementary to the target nucleic acid. In certain embodiments, oligonucleotides are 95% complementary to the target nucleic acid. In certain embodiments, such oligonucleotides are 90%> complementary to the target nucleic acid.

In certain embodiments, such oligonucleotides are 85%> complementary to the target nucleic acid. In certain embodiments, such oligonucleotides are 80%> complementary to the target nucleic acid. In certain embodiments, an antisense compound comprises a region that is fully complementary to a target nucleic acid and is at least 80%> complementary to the target nucleic acid over the entire length of the oligonucleotide. In certain such embodiments, the region of full complementarity is from 6 to 14 nucleobases in length.

Certain Antisense Activities and Mechanisms

In certain antisense activities, hybridization of an antisense compound results in recruitment of a protein that cleaves of the target nucleic acid. For example, certain antisense compounds result in RNase H mediated cleavage of target nucleic acid. RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA:DNA duplex. The "DNA" in such an RNA:DNA duplex, need not be unmodified DNA. In certain embodiments, the invention provides antisense compounds that are sufficiently "DNA-like" to elicit RNase H activity. Such DNA-like antisense compounds include, but are not limited to gapmers having unmodified deoxyfuronose sugar moieties in the nucleosides of the gap and modified sugar moieties in the nucleosides of the wings.

Antisense activities may be observed directly or indirectly. In certain embodiments, observation or detection of an antisense activity involves observation or detection of a change in an amount of a target nucleic acid or protein encoded by such target nucleic acid; a change in the ratio of splice variants of a nucleic acid or protein; and/or a phenotypic change in a cell or animal.

In certain embodiments, compounds comprising oligonucleotides having a gapmer motif described herein have desirable properties compared to non-gapmer oligonucleotides or to gapmers having other motifs. In certain circumstances, it is desirable to identify motifs resulting in a favorable combination of potent antisense activity and relatively low toxicity. In certain embodiments, compounds of the present invention have a favorable therapeutic index (measure of potency divided by measure of toxicity). Certain Target Nucleic Acids

In certain embodiments, antisense compounds comprise or consist of an oligonucleotide comprising a region that is complementary to a target nucleic acid. In certain embodiments, the target nucleic acid is an endogenous RNA molecule. In certain embodiments, the target nucleic acid is a non-coding RNA. In certain such embodiments, the target non-coding RNA is selected from: a long-non-coding RNA, a short non-coding RNA, an intronic RNA molecule, a snoRNA, a scaRNA, a microRNA (including pre-microRNA and mature microRNA), a ribosomal RNA, and promoter directed RNA. In certain embodiments, the target nucleic acid encodes a protein. In certain such embodiments, the target nucleic acid is selected from: an mRNA and a pre-mRNA, including intronic, exonic and untranslated regions. In certain embodiments, oligomeric compounds are at least partially complementary to more than one target nucleic acid. For example, antisense compounds of the present invention may mimic microRNAs, which typically bind to multiple targets.

In certain embodiments, the target nucleic acid is a nucleic acid other than a mature mRNA. In certain embodiments, the target nucleic acid is a nucleic acid other than a mature mRNA or a microRNA. In certain embodiments, the target nucleic acid is a non-coding RNA other than a microRNA. In certain embodiments, the target nucleic acid is a non-coding RNA other than a microRNA or an intronic region of a pre-mRNA. In certain embodiments, the target nucleic acid is a long non-coding RNA. In certain embodiments, the target RNA is an mRNA. In certain embodiments, the target nucleic acid is a pre-mRNA. In certain such embodiments, the target region is entirely within an intron. In certain embodiments, the target region spans an intron/exon junction. In certain embodiments, the target region is at least 50% within an intron. In certain embodiments, the target nucleic acid is selected from among non-coding RNA, including exonic regions of pre-mRNA. In certain embodiments, the target nucleic acid is a ribosomal RNA (rRNA). In certain embodiments, the target nucleic acid is a non-coding RNA associated with splicing of other pre- mRNAs. In certain embodiments, the target nucleic acid is a nuclear-retained non-coding RNA.

In certain embodiments, antisense compounds described herein are complementary to a target nucleic acid comprising a single-nucleotide polymorphism. In certain such embodiments, the antisense compound is capable of modulating expression of one allele of the single-nucleotide polymorphism-containing-target nucleic acid to a greater or lesser extent than it modulates another allele. In certain embodiments an antisense compound hybridizes to a single-nucleotide polymorphism-containing-target nucleic acid at the single- nucleotide polymorphism site. In certain embodiments an antisense compound hybridizes to a single- nucleotide polymorphism-containing-target nucleic acid near the single-nucleotide polymorphism site. In certain embodiments, the target nucleic acid is a Huntingtin gene transcript. In certain embodiments, the target nucleic acid is a single-nucleotide polymorphism-containing-target nucleic acid other than a Huntingtin gene transcript. In certain embodiments, the target nucleic acid is any nucleic acid other than a Huntingtin gene transcript. Certain Pharmaceutical Compositions

In certain embodiments, the present invention provides pharmaceutical compositions comprising one or more antisense compound. In certain embodiments, such pharmaceutical composition comprises a suitable pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutical composition comprises a sterile saline solution and one or more antisense compound. In certain embodiments, such pharmaceutical composition consists of a sterile saline solution and one or more antisense compound. In certain embodiments, the sterile saline is pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition comprises one or more antisense compound and sterile water. In certain embodiments, a pharmaceutical composition consists of one or more antisense compound and sterile water. In certain embodiments, the sterile saline is pharmaceutical grade water. In certain embodiments, a pharmaceutical composition comprises one or more antisense compound and phosphate -buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more antisense compound and sterile phosphate-buffered saline (PBS). In certain embodiments, the sterile saline is pharmaceutical grade PBS.

In certain embodiments, antisense compounds may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations.

Compositions and methods for the formulation of pharmaceutical compositions depend on a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.

Pharmaceutical compositions comprising antisense compounds encompass any pharmaceutically acceptable salts, esters, or salts of such esters. In certain embodiments, pharmaceutical compositions comprising antisense compounds comprise one or more oligonucleotide which, upon administration to an animal, including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of antisense compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.

A prodrug can include the incorporation of additional nucleosides at one or both ends of an oligomeric compound which are cleaved by endogenous nucleases within the body, to form the active antisense oligomeric compound.

Lipid moieties have been used in nucleic acid therapies in a variety of methods. In certain such methods, the nucleic acid is introduced into preformed liposomes or lipoplexes made of mixtures of cationic lipids and neutral lipids. In certain methods, DNA complexes with mono- or poly-cationic lipids are formed without the presence of a neutral lipid. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to a particular cell or tissue. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to fat tissue. In certain embodiments, a lipid moiety is selected to increase distribution of a pharmaceutical agent to muscle tissue. In certain embodiments, pharmaceutical compositions provided herein comprise one or more modified oligonucleotides and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone.

In certain embodiments, a pharmaceutical composition provided herein comprises a delivery system. Examples of delivery systems include, but are not limited to, liposomes and emulsions. Certain delivery systems are useful for preparing certain pharmaceutical compositions including those comprising

hydrophobic compounds. In certain embodiments, certain organic solvents such as dimethylsulfoxide are used.

In certain embodiments, a pharmaceutical composition provided herein comprises one or more tissue- specific delivery molecules designed to deliver the one or more pharmaceutical agents of the present invention to specific tissues or cell types. For example, in certain embodiments, pharmaceutical compositions include liposomes coated with a tissue-specific antibody.

In certain embodiments, a pharmaceutical composition provided herein comprises a co-solvent system. Certain of such co-solvent systems comprise, for example, benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. In certain embodiments, such co-solvent systems are used for hydrophobic compounds. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™ and 65% w/v polyethylene glycol 300. The proportions of such co-solvent systems may be varied considerably without significantly altering their solubility and toxicity characteristics. Furthermore, the identity of co-solvent components may be varied: for example, other surfactants may be used instead of Polysorbate 80™; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.

In certain embodiments, a pharmaceutical composition provided herein is prepared for oral administration. In certain embodiments, pharmaceutical compositions are prepared for buccal administration.

In certain embodiments, a pharmaceutical composition is prepared for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In certain of such embodiments, a pharmaceutical composition comprises a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In certain embodiments, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In certain embodiments, injectable suspensions are prepared using appropriate liquid carriers, suspending agents and the like. Certain pharmaceutical compositions for injection are presented in unit dosage form, e.g., in ampoules or in multi-dose containers. Certain pharmaceutical compositions for injection are suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Certain solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, such suspensions may also contain suitable stabilizers or agents that increase the solubility of the pharmaceutical agents to allow for the preparation of highly concentrated solutions.

In certain embodiments, a pharmaceutical composition is prepared for transmucosal administration. In certain of such embodiments penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

In certain embodiments, a pharmaceutical composition provided herein comprises an oligonucleotide in a therapeutically effective amount. In certain embodiments, the therapeutically effective amount is sufficient to prevent, alleviate or ameliorate symptoms of a disease or to prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art.

In certain embodiments, one or more modified oligonucleotide provided herein is formulated as a prodrug. In certain embodiments, upon in vivo administration, a prodrug is chemically converted to the biologically, pharmaceutically or therapeutically more active form of an oligonucleotide. In certain embodiments, prodrugs are useful because they are easier to administer than the corresponding active form. For example, in certain instances, a prodrug may be more bioavailable (e.g., through oral administration) than is the corresponding active form. In certain instances, a prodrug may have improved solubility compared to the corresponding active form. In certain embodiments, prodrugs are less water soluble than the

corresponding active form. In certain instances, such prodrugs possess superior transmittal across cell membranes, where water solubility is detrimental to mobility. In certain embodiments, a prodrug is an ester. In certain such embodiments, the ester is metabolically hydrolyzed to carboxylic acid upon administration. In certain instances the carboxylic acid containing compound is the corresponding active form. In certain embodiments, a prodrug comprises a short peptide (polyaminoacid) bound to an acid group. In certain of such embodiments, the peptide is cleaved upon administration to form the corresponding active form.

In certain embodiments, the present invention provides compositions and methods for reducing the amount or activity of a target nucleic acid in a cell. In certain embodiments, the cell is in an animal. In certain embodiments, the animal is a mammal. In certain embodiments, the animal is a rodent. In certain embodiments, the animal is a primate. In certain embodiments, the animal is a non-human primate. In certain embodiments, the animal is a human.

In certain embodiments, the present invention provides methods of administering a pharmaceutical composition comprising an oligomeric compound of the present invention to an animal. Suitable administration routes include, but are not limited to, oral, rectal, transmucosal, intestinal, enteral, topical, suppository, through inhalation, intrathecal, intracerebroventricular, intraperitoneal, intranasal, intraocular, intratumoral, and parenteral (e.g., intravenous, intramuscular, intramedullary, and subcutaneous). In certain embodiments, pharmaceutical intrathecals are administered to achieve local rather than systemic exposures.

Nonlimiting disclosure and incorporation by reference

While certain compounds, compositions and methods described herein have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds described herein and are not intended to limit the same. Each of the references, GenBank accession numbers, and the like recited in the present application is incorporated herein by reference in its entirety.

Although the sequence listing accompanying this filing identifies each sequence as either "RNA" or "DNA" as required, in reality, those sequences may be modified with any combination of chemical modifications. One of skill in the art will readily appreciate that such designation as "RNA" or "DNA" to describe modified oligonucleotides is, in certain instances, arbitrary. For example, an oligonucleotide comprising a nucleoside comprising a 2' -OH sugar moiety and a thymine base could be described as a DNA having a modified sugar (2'-OH for the natural 2'-H of DNA) or as an RNA having a modified base (thymine (methylated uracil) for natural uracil of RNA).

Accordingly, nucleic acid sequences provided herein, including, but not limited to those in the sequence listing, are intended to encompass nucleic acids containing any combination of natural or modified RNA and/or DNA, including, but not limited to such nucleic acids having modified nucleobases. By way of further example and without limitation, an oligomeric compound having the nucleobase sequence

"ATCGATCG" encompasses any oligomeric compounds having such nucleobase sequence, whether modified or unmodified, including, but not limited to, such compounds comprising RNA bases, such as those having sequence "AUCGAUCG" and those having some DNA bases and some RNA bases such as

"AUCGATCG" and oligomeric compounds having other modified or naturally occurring bases, such as "ATmeCGAUCG," wherein meC indicates a cytosine base comprising a methyl group at the 5-position.

EXAMPLES

The following examples illustrate certain embodiments of the present invention and are not limiting. Moreover, where specific embodiments are provided, the inventors have contemplated generic application of those specific embodiments. For example, disclosure of an oligonucleotide having a particular motif provides reasonable support for additional oligonucleotides having the same or similar motif. And, for example, where a particular high-affinity modification appears at a particular position, other high-affinity modifications at the same position are considered suitable, unless otherwise indicated.

Where nucleobase sequences are not provided, to allow assessment of the relative effects of nucleobase sequence and chemical modification, throughout the examples, oligomeric compounds are assigned a "Sequence Code." Oligomeric compounds having the same Sequence Code have the same nucleobase sequence. Oligomeric compounds having different Sequence Codes have different nucleobase sequences.

Example 1: Dose-dependent inhibition of chimeric antisense oligonucleotides targeting PTEN

A series of modified oligonucleotides were designed based on the parent gapmer, ISIS 482050, wherein the central gap region contains ten 2'-deoxynucleosides. These modified oligonucleotides were designed by having the central gap region shortened to nine, eight or seven 2'-deoxynucleosides and by introducing 2'-0-methoxyethyl (MOE) modifications at one or both wing regions. The newly designed oligonucleotides were evaluated for their effecst in reducing PTEN mRNA levels in vitro.

The gapmers and their motifs are described in Table 60. The internucleoside linkages throughout each gapmer are phosphorothioate linkages (P=S). Nucleosides followed by a subscript "d" indicate 2'- deoxynucleosides. Nucleosides followed by a subscript "e" indicate 2'-0-methoxyethyl (MOE) nucleosides. Nucleosides followed by a subscript "k" indicate constrained ethyl (cEt) nucleosides. "N" indicates modified or naturally occurring nucleobases (A, T, C, G, U, or 5-methyl C).

The newly designed gapmers were tested in vitro. Mouse primary hepatocytes were plated at a density of 20,000 cells per well and transfected using electroporation with 0.6 μΜ, 3.0 μΜ and 15 μΜ concentrations of antisense oligonucleotides. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PTEN mRNA levels were measured by quantitative real-time PCR. Mouse PTEN primer probe set RTS186 was used to measure mRNA levels. PTEN mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. The results in Table 12 are presented as PTEN mRNA expression relative to untreated control cells (% UTC).

The parent gapmer, ISIS 482050 was included in the study as a bench mark oligonucleotide against which the activity of the newly designed gapmers targeting PTEN could be compared.

Table 11

Chimeric antisense oligonucleotides targeting PTEN

Figure imgf000112_0001
dAdGk mCkTeTe

AeTk mCkAdTdGdGd mCdTdGd mC

573356 3-8-5 Full deoxy ekk kkeee 23

dAkGk CeTeTe

AkTk mCkAdTdGdGd mCdTdGd mC

573357 3-7-6 Full deoxy ekk kkeeee 23

kAkGe CeTeTe

AeTe mCkAkTdGdGd mCdTdGd mC

573358 4-8-4 Full deoxy eekk kkee 23

dAdGk mCkTeTe

AeTe mCeAkTkGdGd mCdTdGd mC

573359 5-7-4 Full deoxy eeekk kkee 23

dAdGk CkTeTe

AeTe mCkAkTdGdGd mCdTdGd mC

573360 4-7-5 Full deoxy eekk kkeee 23

dAkGk mCeTeTe

e = 2' -MOE, k = cEt, d= 2'-deoxynucleoside

Table 12

Dose-response effect of chimeric antisense oligonucleotides targeting PTEN

Figure imgf000113_0001

e = 2' -MOE, k = cEt, d= 2'-deoxynucleoside

Example 2: Dose-dependent inhibition of chimeric antisense oligonucleotides targeting PTEN

Additional chimeric oligonucleotides were designed based on the parent gapmer, ISIS 482050, wherein the central gap region contains ten 2'-deoxynucleosides. These modified oligonucleotides were designed by having the central gap region shortened to eight 2'-deoxynucleosides and by introducing one or more 2'-0-methoxyethyl (MOE) modification(s) at the 3' wing region. The modified oligonucleotides designed by microwalk were evaluated for their effects in reducing PTEN mRNA levels in vitro.

The gapmers and their motifs are described in Table 13. The internucleoside linkages throughout each gapmer are phosphorothioate linkages (P=S). Nucleosides followed by a subscript "d" indicate 2'- deoxynucleoside. Nucleosides followed by a subscript "e" indicate 2'-0-methoxyethyl (MOE) nucleosides. Nucleosides followed by a subscript "k" indicate constrained ethyl (cEt) nucleosides. mC indicates a 5- methyl nucleoside. The newly designed gapmers were tested in vitro. Mouse primary hepatocytes were plated at a density of 20,000 cells per well and transfected using electroporation with 0.6 μΜ, 3.0 μΜ and 15 μΜ concentrations of antisense oligonucleotides. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PTEN mRNA levels were measured by quantitative real-time PCR. Mouse PTEN primer probe set RTS186 was used to measure mRNA levels. PTEN mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. The results in Table 14 are presented as PTEN mRNA expression relative to untreated control cells (% UTC).

The parent gapmer, ISIS 482050 was included in the study as a bench mark oligonucleotide against which the activity of the newly designed gapmers targeting PTEN could be compared.

Table 13

Chimeric antisense oligonucleotides designed by microwalk targeting PTEN

Figure imgf000114_0001

e = 2'-MOE, k = cEt, d= 2'-deoxynucleoside

Table 14

Dose-dependent inhibition of chimeric antisense oligonucleotides designed by microwalk targeting PTEN

Figure imgf000114_0002
573800 68.6 38.9 12.0 3-8-5 Full deoxy kkk keeee

573801 54.6 46.3 11.8 3-8-5 Full deoxy kkk keeee

573802 60.7 40.4 13.0 3-8-5 Full deoxy kkk keeee

573803 47.0 29.8 8.5 3-8-5 Full deoxy kkk keeee

573804 62.5 34.1 11.3 3-8-5 Full deoxy kkk keeee

573805 70.3 31.6 15.2 3-8-5 Full deoxy kkk keeee

Saline = 100

e = 2'-MOE, k = cEt, d= 2'-deoxynucleoside

Example 3: Antisense inhibition of Target-Z mRNA in HepG2 cells

Antisense oligonucleotides were designed targeting a Target-Z nucleic acid and were tested for their effects on Target-Z mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables shown below. ISIS 146786, 509934, ISIS 509959, and ISIS 510100, were also included in these studies for comparison. Cultured HepG2 cells at a density of 28,000 cells per well were transfected using

LipofectAMINE2000® with 70 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Target-Z mRNA levels were measured by quantitative real-time PCR. Viral primer probe set RTS3370 (forward sequence CTTGGTCATGGGCCATCAG, designated herein as SEQ ID NO: 33; reverse sequence CGGCTAGGAGTTCCGCAGTA, designated herein as SEQ ID NO: 34; probe sequence TGCGTGGAACCTTTTCGGCTCC, designated herein as SEQ ID NO: 35) was used to measure mRNA levels. Levels were also measured using primer probe set RTS3371 (forward sequence CCAAACCTTCGGACGGAAA, designated herein as SEQ ID NO: 36; reverse sequence

TGAGGCCCACTCCCATAGG, designated herein as SEQ ID NO: 37; probe sequence

CCCATCATCCTGGGCTTTCGGAAAAT, designated herein as SEQ ID NO: 38). Target-Z mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target-Z, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides and their motifs are described in Tables 15- 20. The gapmers are 16 nucleotides in length, wherein the central gap region comprises ten 2'- deoxynucleosides. Nucleosides followed by 'k' indicate constrained ethyl (cEt) nucleosides. Nucleosides followed by "e" indicate 2'-0-methoxyethyl (2'-MOE) nucleosides. The internucleoside linkages throughout each gapmer are phosphorothioate (P=S) linkages. All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

Each gapmer listed in Tables 15-20 is targeted to the viral genomic sequence, designated herein as Target-Z. The activity of the newly designed oligonucleotides was compared with ISIS 146786, ISIS 509934, ISIS 509959, and ISIS 510100. Table 15

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000116_0001
552825 ekk-d(10)-kke 51

552826 ekk-d(10)-kke 55

552827 ekk-d(10)-kke 67

552828 ekk-d(10)-kke 78

552829 ekk-d(10)-kke 72

552830 ekk-d(10)-kke 71

552831 ekk-d(10)-kke 69

552832 ekk-d(10)-kke 67

552833 ekk-d(10)-kke 65

552834 ekk-d(10)-kke 78

552835 ekk-d(10)-kke 70

552836 ekk-d(10)-kke 64

552837 ekk-d(10)-kke 65

552838 ekk-d(10)-kke 64

552839 ekk-d(10)-kke 60

552840 ekk-d(10)-kke 35

552841 ekk-d(10)-kke 62

552842 ekk-d(10)-kke 67

552843 ekk-d(10)-kke 77

552844 ekk-d(10)-kke 81

552845 ekk-d(10)-kke 63

552846 ekk-d(10)-kke 79

552847 ekk-d(10)-kke 47

552848 ekk-d(10)-kke 69

552849 ekk-d(10)-kke 59

552850 ekk-d(10)-kke 83

552851 ekk-d(10)-kke 90

552852 ekk-d(10)-kke 89

552853 ekk-d(10)-kke 83

552854 ekk-d(10)-kke 80

552855 ekk-d(10)-kke 75

552856 ekk-d(10)-kke 69

552857 ekk-d(10)-kke 68

552858 ekk-d(10)-kke 79

552859 ekk-d(10)-kke 79

552860 ekk-d(10)-kke 71

552861 ekk-d(10)-kke 68

552862 ekk-d(10)-kke 65

552863 ekk-d(10)-kke 70

552864 ekk-d(10)-kke 71

e = 2'-MOE, k = cEt, d = 2'-deoxynucleoside

Table 16

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371 %

ISIS No Motif

inhibition

552787 ekk-d(10)-kke 53

552788 ekk-d(10)-kke 45

552789 ekk-d(10)-kke 75

552790 ekk-d(10)-kke 68

552791 ekk-d(10)-kke 51

552792 ekk-d(10)-kke 38

552793 ekk-d(10)-kke 0

552794 ekk-d(10)-kke 44

552795 ekk-d(10)-kke 56

552796 ekk-d(10)-kke 45

552797 ekk-d(10)-kke 46

552798 ekk-d(10)-kke 53

552799 ekk-d(10)-kke 48

552800 ekk-d(10)-kke 54

552801 ekk-d(10)-kke 63

552802 ekk-d(10)-kke 49

552803 ekk-d(10)-kke 71

552804 ekk-d(10)-kke 64

552805 ekk-d(10)-kke 70

552806 ekk-d(10)-kke 67

552807 ekk-d(10)-kke 61

552808 ekk-d(10)-kke 83

552809 ekk-d(10)-kke 59

552810 ekk-d(10)-kke 56

55281 1 ekk-d(10)-kke 62

552812 ekk-d(10)-kke 66

552813 ekk-d(10)-kke 63

552814 ekk-d(10)-kke 65

552815 ekk-d(10)-kke 63

552816 ekk-d(10)-kke 88

552817 ekk-d(10)-kke 94

552818 ekk-d(10)-kke 82

552819 ekk-d(10)-kke 80

552820 ekk-d(10)-kke 84

552821 ekk-d(10)-kke 71

552822 ekk-d(10)-kke 85

552823 ekk-d(10)-kke 71

552824 ekk-d(10)-kke 81

552825 ekk-d(10)-kke 51

552826 ekk-d(10)-kke 64

552827 ekk-d(10)-kke 61 552828 ekk-d(10)-kke 76

552829 ekk-d(10)-kke 61

552830 ekk-d(10)-kke 59

552831 ekk-d(10)-kke 58

552832 ekk-d(10)-kke 64

552833 ekk-d(10)-kke 75

552834 ekk-d(10)-kke 84

552835 ekk-d(10)-kke 57

552836 ekk-d(10)-kke 51

552837 ekk-d(10)-kke 53

552838 ekk-d(10)-kke 48

552839 ekk-d(10)-kke 50

552840 ekk-d(10)-kke 54

552841 ekk-d(10)-kke 61

552842 ekk-d(10)-kke 71

552843 ekk-d(10)-kke 75

552844 ekk-d(10)-kke 78

552845 ekk-d(10)-kke 52

552846 ekk-d(10)-kke 76

552847 ekk-d(10)-kke 61

552848 ekk-d(10)-kke 72

552849 ekk-d(10)-kke 87

552850 ekk-d(10)-kke 76

552851 ekk-d(10)-kke 76

552852 ekk-d(10)-kke 79

552853 ekk-d(10)-kke 82

552854 ekk-d(10)-kke 85

552855 ekk-d(10)-kke 78

552856 ekk-d(10)-kke 77

552857 ekk-d(10)-kke 75

552858 ekk-d(10)-kke 75

552859 ekk-d(10)-kke 79

552860 ekk-d(10)-kke 71

552861 ekk-d(10)-kke 74

552862 ekk-d(10)-kke 66

552863 ekk-d(10)-kke 70

552864 ekk-d(10)-kke 73 e = 2'-MOE, k = cEt, d = 2'-deoxynucleoside Table 17

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000120_0001
552928 ek-d(10)-keke 78

552929 ek-d(10)-keke 75

552930 ek-d(10)-keke 78

552931 ek-d(10)-keke 74

552932 ek-d(10)-keke 86

552933 ek-d(10)-keke 82

552934 ek-d(10)-keke 74

552935 ek-d(10)-keke 76

552936 ek-d(10)-keke 81

552937 ek-d(10)-keke 80

552938 ek-d(10)-keke 78

552939 ek-d(10)-keke 75

552940 ek-d(10)-keke 63

552941 ekk-d(10)-kke 78

552942 ek-d(10)-keke 80

552865 ekk-d(10)-kke 67

552866 ekk-d(10)-kke 68

552868 ekk-d(10)-kke 55

552869 ekk-d(10)-kke 48

552870 ekk-d(10)-kke 55

552871 ekk-d(10)-kke 57

552872 ekk-d(10)-kke 70

552873 ekk-d(10)-kke 49

552874 ekk-d(10)-kke 42

552875 ekk-d(10)-kke 41

552876 ekk-d(10)-kke 50

552877 ek-d(10)-keke 39

552878 ekk-d(10)-kke 31

552879 ekk-d(10)-kke 5

552880 ekk-d(10)-kke 5

552881 ekk-d(10)-kke 10

552882 ekk-d(10)-kke 1 1

552883 ekk-d(10)-kke 27

552884 ekk-d(10)-kke 36

552885 ekk-d(10)-kke 12

552886 ekk-d(10)-kke 32

552888 ekk-d(10)-kke 1 e = 2'-MOE, k = cEt, d = 2'-deoxynucleoside Table 18

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000122_0001

21 552993 eee-d(10)-kkk 52

552994 eee-d(10)-kkk 58

552995 eee-d(10)-kkk 70

552996 eee-d(10)-kkk 74

552997 eee-d(10)-kkk 59

552998 eee-d(10)-kkk 82

552999 eee-d(10)-kkk 70

553000 eee-d(10)-kkk 67

553001 eee-d(10)-kkk 67

553002 eee-d(10)-kkk 74

553003 eee-d(10)-kkk 72

553004 eee-d(10)-kkk 73

553005 eee-d(10)-kkk 67

553006 eee-d(10)-kkk 69

553007 eee-d(10)-kkk 60

553008 eee-d(10)-kkk 71

552943 ek-d(10)-keke 77

553009 eee-d(10)-kkk 78

552944 ek-d(10)-keke 74

553010 eee-d(10)-kkk 78

552945 ek-d(10)-keke 76

55301 1 eee-d(10)-kkk 72

552946 ek-d(10)-keke 71

553012 eee-d(10)-kkk 74

552947 ek-d(10)-keke 54

553013 eee-d(10)-kkk 39

552948 ek-d(10)-keke 50

553014 eee-d(10)-kkk 37

552949 ek-d(10)-keke 8

553015 eee-d(10)-kkk 45

552950 ek-d(10)-keke 44

553016 eee-d(10)-kkk 47

552951 ek-d(10)-keke 60

553017 eee-d(10)-kkk 47

552952 ek-d(10)-keke 35

553018 eee-d(10)-kkk 30

552953 ek-d(10)-keke 37

553019 eee-d(10)-kkk 37

552954 ek-d(10)-keke 40

553020 eee-d(10)-kkk 24 e = 2'-MOE, k = cEt, d = 2'-deoxynucleoside Table 19

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000124_0001
552929 ek-d(10)-keke 68

552930 ek-d(10)-keke 87

552931 ek-d(10)-keke 87

552932 ek-d(10)-keke 88

552933 ek-d(10)-keke 75

552934 ek-d(10)-keke 76

552935 ek-d(10)-keke 71

552936 ek-d(10)-keke 80

552937 ek-d(10)-keke 81

552938 ek-d(10)-keke 85

552939 ek-d(10)-keke 82

552940 ek-d(10)-keke 76

552941 ekk-d(10)-kke 72

552942 ek-d(10)-keke 85

552865 ekk-d(10)-kke 70

552866 ekk-d(10)-kke 65

552868 ekk-d(10)-kke 36

552869 ekk-d(10)-kke 23

552870 ekk-d(10)-kke 49

552871 ekk-d(10)-kke 46

552872 ekk-d(10)-kke 73

552873 ekk-d(10)-kke 41

552874 ekk-d(10)-kke 18

552875 ekk-d(10)-kke 0

552876 ekk-d(10)-kke 49

552877 ek-d(10)-keke 37

552878 ekk-d(10)-kke 28

552879 ekk-d(10)-kke 0

552880 ekk-d(10)-kke 12

552881 ekk-d(10)-kke 0

552882 ekk-d(10)-kke 0

552883 ekk-d(10)-kke 12

552884 ekk-d(10)-kke 39

552885 ekk-d(10)-kke 37

552886 ekk-d(10)-kke 15

552888 ekk-d(10)-kke 0

e = 2'-MOE, k = cEt, d = 2'-deoxynucleoside

Table 20

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000125_0001
552957 eee-d(10)-kkk 73

552958 eee-d(10)-kkk 63

552959 eee-d(10)-kkk 58

552960 eee-d(10)-kkk 67

552961 eee-d(10)-kkk 78

552962 eee-d(10)-kkk 29

552963 eee-d(10)-kkk 25

552964 eee-d(10)-kkk 33

552965 eee-d(10)-kkk 55

552966 eee-d(10)-kkk 71

552967 eee-d(10)-kkk 23

552968 eee-d(10)-kkk 41

552969 eee-d(10)-kkk 76

552970 eee-d(10)-kkk 44

552971 eee-d(10)-kkk 77

552972 eee-d(10)-kkk 74

552973 eee-d(10)-kkk 61

552974 eee-d(10)-kkk 73

552975 eee-d(10)-kkk 66

552976 eee-d(10)-kkk 70

552977 eee-d(10)-kkk 65

552978 eee-d(10)-kkk 40

552979 eee-d(10)-kkk 79

552980 eee-d(10)-kkk 81

552981 eee-d(10)-kkk 74

552982 ek-d(10)-keke 52

552983 eee-d(10)-kkk 78

552984 eee-d(10)-kkk 71

552985 eee-d(10)-kkk 38

552986 eee-d(10)-kkk 48

552987 eee-d(10)-kkk 54

552988 eee-d(10)-kkk 85

552989 eee-d(10)-kkk 84

552990 eee-d(10)-kkk 79

552991 eee-d(10)-kkk 53

552992 eee-d(10)-kkk 68

552993 eee-d(10)-kkk 67

552994 eee-d(10)-kkk 69

552995 eee-d(10)-kkk 62

552996 eee-d(10)-kkk 82

552997 eee-d(10)-kkk 58

552998 eee-d(10)-kkk 86

552999 eee-d(10)-kkk 63 553000 eee-d(10)-kkk 67

553001 eee-d(10)-kkk 70

553002 eee-d(10)-kkk 84

553003 eee-d(10)-kkk 83

553004 eee-d(10)-kkk 68

553005 eee-d(10)-kkk 57

553006 eee-d(10)-kkk 74

553007 eee-d(10)-kkk 62

553008 eee-d(10)-kkk 50

552943 ek-d(10)-keke 86

553009 eee-d(10)-kkk 79

552944 ek-d(10)-keke 83

553010 eee-d(10)-kkk 74

552945 ek-d(10)-keke 79

55301 1 eee-d(10)-kkk 60

552946 ek-d(10)-keke 68

553012 eee-d(10)-kkk 78

552947 ek-d(10)-keke 51

553013 eee-d(10)-kkk 45

552948 ek-d(10)-keke 56

553014 eee-d(10)-kkk 53

552949 ek-d(10)-keke 1

553015 eee-d(10)-kkk 55

552950 ek-d(10)-keke 52

553016 eee-d(10)-kkk 65

552951 ek-d(10)-keke 59

553017 eee-d(10)-kkk 36

552952 ek-d(10)-keke 34

553018 eee-d(10)-kkk 20

552953 ek-d(10)-keke 55

553019 eee-d(10)-kkk 34

552954 ek-d(10)-keke 51

553020 eee-d(10)-kkk 28

e = 2'-MOE, k = cE , d = 2'-deoxynucleoside

Example 4: Dose-dependent antisense inhibition of Target-Z mRNA in HepG2 cells

Antisense oligonucleotides from the study described in Example 46 exhibiting in vitro inhibition of Target-Z mRNA were selected and tested at various doses in HepG2 cells. Cells were plated at a density of 28,000 cells per well and transfected using LipofectAMINE2000® with 9.26 nM, 27.78 nM, 83.33 nM, and 250.00 nM concentrations of antisense oligonucleotide, as specified in Table 21. After a treatment period of approximately 16 hours, RNA was isolated from the cells and Target-Z mRNA levels were measured by quantitative real-time PCR. Target-Z primer probe set RTS3371 was used to measure mRNA levels. Target- Z mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target-Z, relative to untreated control cells.

As illustrated in Table 21, Target-Z mRNA levels were reduced in a dose-dependent manner in antisense oligonucleotide treated cells.

Table 21

Dose-dependent antisense inhibition of human Target-Z in HepG2 cells

Figure imgf000128_0001
552998 eee-d(10)-kkk 17 49 79 90

553002 eee-d(10)-kkk 0 32 68 86

553003 eee-d(10)-kkk 15 42 67 88 e = 2' -MOE, k = cEt, d = 2'-deoxynucleoside

Example 5: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Mice harboring a Target-Z gene fragment (Guidotti, L. G. et al., J. Virol. 1995, 69, 6158-6169) were used. The mice were treated with ISIS antisense oligonucleotides selected from studies described above as illustrated in Table 22 and evaluated for their efficacy in this model.

Treatment

Groups of 10 mice each were injected subcutaneously twice a week for the first with 50 mg/kg and, subsequently, twice a week for the next 3 weeks with 25 mg/kg of ISIS 146786 or ISIS 510100. Control groups of 10 mice each were treated in a similar manner with ISIS 141923 (5-10-5 MOE gapmer with no known murine target) or ISIS 459024 (3-10-4 MOE gapmer with no known murine target). Mice were euthanized 48 hours after the last dose, and organs and serum were harvested for further analysis.

Table 22

Antisense oligonucleotides targeting Target-Z in transgenic mice

Figure imgf000129_0001

e = 2'-MOE (e.g. e5 = eeeee), d = 2'-deoxynucleoside

DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe sets RTS3370, RTS3371, or RTS3372 (forward sequence ATCCTATCAACACTTCCGGAAACT, designated SEQ ID NO: 43; reverse sequence CGACGCGGCGATTGAG, designated SEQ ID NO: 44; probe sequence AAGAACTCCCTCGCCTCGCAGACG, designated SEQ ID NO: 45). The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe sets RTS3370 and RTS3371 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. The data is presented in Table 23. Serum DNA samples were analyzed after the study period. The data is presented in Table 24, expressed relative to the levels measured in the control group. As shown in Tables 23 and 24, the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control. Treatment with either control oligonucleotide did not cause any changes in RNA or DNA levels, as expected.

Table 23

Percent inhibition of Target-Z RNA and DNA in the liver of transgenic mice

Figure imgf000130_0001

e = 2'-MOE (e.g. e5 = eeeee), d = 2'-deoxynucleoside

Table 24

Percent inhibition of Target-Z DNA in the serum of transgenic mice

Figure imgf000130_0002

e = 2'-MOE (e.g. e5 = eeeee), d = 2'-deoxynucleoside

Example 6: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Transgenic mice were treated with ISIS antisense oligonucleotides selected from studies described above and evaluated for their efficacy in this model.

Treatment

A group of 6 mice was injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786. Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 10 mg/kg of ISIS 552803, ISIS 552903, ISIS 552817, ISIS 552822, and ISIS 552907. One group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe set RTS3371. The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe set RTS3371 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. The data is presented in Table 25. Serum DNA samples were analyzed after the study period. The data is presented in Table 26, expressed relative to the levels measured in the control group. As shown in Tables 25 and 26, the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control.

Table 25

Percent inhibition of Target-Z RNA and DNA in transgenic mice

Figure imgf000131_0001

e = 2'-MOE (e.g. e5 = eeeee), d = 2'-deoxynucleoside

Table 26

Serum levels of Target-Z DNA in transgenic mice, relative to control levels

Figure imgf000131_0002

e = 2'-MOE (e.g. e5 = eeeee), d = 2'-deoxynucleoside

Liver function

To evaluate the effect of ISIS oligonucleotides on hepatic function, plasma concentrations of ALT were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY) (Nyblom, H. et al., Alcohol & Alcoholism 39: 336-339, 2004; Tietz NW (Ed): Clinical Guide to Laboratory Tests, 3rd ed. W. B. Saunders, Philadelphia, PA, 1995). The results are presented in Table 27 expressed in IU/L. All the ISIS oligonucleotides were considered tolerable in the mice, as demonstrated by their liver transaminase profile. Table 27

ALT levels (IU/L) of transgenic mice

Figure imgf000132_0001

e = 2'-MOE (e.g. e5 = eeeee), d = 2'-deoxynucleoside

Example 7: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Transgenic mice were treated with ISIS antisense oligonucleotides selected from studies described above and evaluated for their efficacy in this model.

Treatment

A group of 6 mice was injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786. Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 10 mg/kg of ISIS 552853, ISIS 552854, ISIS 552932, and ISIS 552938. One group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe set RTS3371. The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe set RTS3371 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. As shown in Table 28, the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control. Results are presented as percent inhibition of Target-Z mRNA or DNA, relative to control.

Table 28

Percent inhibition of Target-Z DNA and RNA in transgenic mice

Figure imgf000132_0002
ISIS 552932 ekk-d(10)-kke 20 75 70

ISIS 552938 ek-d(10)-keke 20 67 56

= 2'-M0E (e.g. e5 = eeeee), d = 2'-deoxynucleoside

Example 8: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Transgenic mice were treated with ISIS antisense oligonucleotides selected from studies described above and evaluated for their efficacy in this model.

Treatment

A group of 6 mice was injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786. Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 10 mg/kg of ISIS 552922, ISIS 552923, ISIS 552942, ISIS 552872, ISIS 552925, ISIS 552937, and ISIS 552939. One group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe set RTS3371. The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe set RTS3371 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. As shown in Table 29, the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control. Results are presented as percent inhibition of Target-Z mRNA or DNA, relative to control.

Table 29

Percent inhibition of Target-Z DNA and RNA in transgenic mice

Figure imgf000133_0001

= 2'-MOE (e.g. e5 = eeeee), d = 2'-deoxynucleoside Example 9: Antisense inhibition of Target-Z mRNA in HepG2 cells

Antisense oligonucleotides were designed targeting a Target-Z nucleic acid and were tested for their effects on Target-Z mRNA in vitro. The antisense oligonucleotides were tested in a series of experiments that had similar culture conditions. The results for each experiment are presented in separate tables. ISIS 146786, 509934, ISIS 509959, and ISIS 510100, from the studies described above, were also included. Cultured HepG2 cells at a density of 28,000 cells per well were transfected using LipofectAMINE2000® with 70 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Target-Z mRNA levels were measured by quantitative real-time PCR. Primer probe set RTS3370 (forward sequence CTTGGTCATGGGCCATCAG, designated herein as SEQ ID NO: 33; reverse sequence CGGCTAGGAGTTCCGCAGTA, designated herein as SEQ ID NO: 34; probe sequence TGCGTGGAACCTTTTCGGCTCC, designated herein as SEQ ID NO: 35) was used to measure mRNA levels. Levels were also measured using primer probe set RTS3371 (forward sequence

CCAAACCTTCGGACGGAAA, designated herein as SEQ ID NO: 36; reverse sequence

TGAGGCCCACTCCCATAGG, designated herein as SEQ ID NO: 37; probe sequence

CCCATCATCCTGGGCTTTCGGAAAAT, designated herein as SEQ ID NO: 38). Target-Z mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target-Z, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides and their motifs are described in Tables 30- 47. The modified oligonucleotides are 16, 17 or 20 nucleotides in length, wherein the central gap segment comprises of nine or ten 2'-deoxynucleosides and is flanked by wing segments on the 5' direction and the 3' direction comprising 2'-0-methoxyethyl (2'-MOE) modifications. The internucleoside linkages throughout each gapmer are phosphorothioate (P=S) linkages. All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

Each gapmer listed in the Tables is targeted to the viral genomic sequence, designated herein as Target-Z. The activity of the newly designed oligonucleotides was compared with ISIS 146786, 509934, ISIS 509959, and ISIS 510100, the information of which have been placed at the top of each table.

Table 30

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000134_0001
552279 5-9-3 2'-MOE 41

552280 5-9-3 2'-MOE 5

552281 5-9-3 2'-MOE 1 1

552282 5-9-3 2'-MOE 20

552283 5-9-3 2'-MOE 28

552230 4-9-4 2'-MOE 57

552284 5-9-3 2'-MOE 0

552231 4-9-4 2'-MOE 29

552285 5-9-3 2'-MOE 61

552232 4-9-4 2'-MOE 35

552286 5-9-3 2'-MOE 47

552233 4-9-4 2'-MOE 38

552287 5-9-3 2'-MOE 45

552234 4-9-4 2'-MOE 0

552288 5-9-3 2'-MOE 50

552235 4-9-4 2'-MOE 0

552289 5-9-3 2'-MOE 46

552236 4-9-4 2'-MOE 45

552290 5-9-3 2'-MOE 41

552237 4-9-4 2'-MOE 44

552291 5-9-3 2'-MOE 26

552239 4-9-4 2'-MOE 62

552293 5-9-3 2'-MOE 67

552240 4-9-4 2'-MOE 61

552294 5-9-3 2'-MOE 71

552241 4-9-4 2'-MOE 55

552295 5-9-3 2'-MOE 58

552242 4-9-4 2'-MOE 60

552296 5-9-3 2'-MOE 59

552243 4-9-4 2'-MOE 57

552297 5-9-3 2'-MOE 55

552244 4-9-4 2'-MOE 33

552298 5-9-3 2'-MOE 48

552245 4-9-4 2'-MOE 48

552299 5-9-3 2'-MOE 34

552246 4-9-4 2'-MOE 81

552300 5-9-3 2'-MOE 56

552247 4-9-4 2'-MOE 87

552301 5-9-3 2'-MOE 86

552248 4-9-4 2'-MOE 72

552302 5-9-3 2'-MOE 77

552249 4-9-4 2'-MOE 56

552303 5-9-3 2'-MOE 65 552250 4-9-4 2'-MOE 52

552304 5-9-3 2'-MOE 57

552251 4-9-4 2'-MOE 43

552305 5-9-3 2'-MOE 56

552252 4-9-4 2'-MOE 62

552306 5-9-3 2'-MOE 75

552253 4-9-4 2'-MOE 82

552307 5-9-3 2'-MOE 90

552254 4-9-4 2'-MOE 74

552255 4-9-4 2'-MOE 78

552256 4-9-4 2'-MOE 65

552257 4-9-4 2'-MOE 62

552258 4-9-4 2'-MOE 72

552259 4-9-4 2'-MOE 63

552260 4-9-4 2'-MOE 58

552261 4-9-4 2'-MOE 63

552262 4-9-4 2'-MOE 50

552263 4-9-4 2'-MOE 60

552264 4-9-4 2'-MOE 52

552265 4-9-4 2'-MOE 68

552266 4-9-4 2'-MOE 62

552267 4-9-4 2'-MOE 58

552268 4-9-4 2'-MOE 62

552269 4-9-4 2'-MOE 52

552270 4-9-4 2'-MOE 54

552271 4-9-4 2'-MOE 58

552272 4-9-4 2'-MOE 40

552273 4-9-4 2'-MOE 34

552274 4-9-4 2'-MOE 34

552275 4-9-4 2'-MOE 39

Table 31

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000136_0001
552442 3-9-4 2'-MOE 53

552387 2-9-5 2'-MOE 48

552443 3-9-4 2'-MOE 59

552388 2-9-5 2'-MOE 40

552444 3-9-4 2'-MOE 51

552389 2-9-5 2'-MOE 39

552445 3-9-4 2'-MOE 60

552390 2-9-5 2'-MOE 52

552446 3-9-4 2'-MOE 54

552391 2-9-5 2'-MOE 57

552447 3-9-4 2'-MOE 54

552392 2-9-5 2'-MOE 0

552448 3-9-4 2'-MOE 58

552393 2-9-5 2'-MOE 59

552449 3-9-4 2'-MOE 60

552394 2-9-5 2'-MOE 53

552450 3-9-4 2'-MOE 53

552395 2-9-5 2'-MOE 57

552451 3-9-4 2'-MOE 39

552396 2-9-5 2'-MOE 62

552452 3-9-4 2'-MOE 57

552238 4-9-4 2'-MOE 38

552292 5-9-3 2'-MOE 48

552346 6-9-2 2'-MOE 0

552397 2-9-5 2'-MOE 63

552453 3-9-4 2'-MOE 56

552398 2-9-5 2'-MOE 61

552454 3-9-4 2'-MOE 48

552399 2-9-5 2'-MOE 52

552400 2-9-5 2'-MOE 57

552401 2-9-5 2'-MOE 52

552402 2-9-5 2'-MOE 54

552403 2-9-5 2'-MOE 74

552404 2-9-5 2'-MOE 43

552405 2-9-5 2'-MOE 15

552406 2-9-5 2'-MOE 37

552407 2-9-5 2'-MOE 37

552408 2-9-5 2'-MOE 76

552409 2-9-5 2'-MOE 76

552410 2-9-5 2'-MOE 63

55241 1 2-9-5 2'-MOE 70

552412 2-9-5 2'-MOE 62

552413 2-9-5 2'-MOE 56

36 552414 2-9-5 2'-MOE 63

552415 2-9-5 2'-MOE 52

552416 2-9-5 2'-MOE 67

552417 2-9-5 2'-MOE 50

552418 2-9-5 2'-MOE 79

552419 2-9-5 2'-MOE 70

552420 2-9-5 2'-MOE 71

552421 2-9-5 2'-MOE 69

552422 2-9-5 2'-MOE 68

552423 2-9-5 2'-MOE 65

552424 2-9-5 2'-MOE 70

552425 2-9-5 2'-MOE 51

552426 2-9-5 2'-MOE 40

552427 2-9-5 2'-MOE 35

552428 2-9-5 2'-MOE 58

552429 2-9-5 2'-MOE 46

552430 2-9-5 2'-MOE 53

552431 2-9-5 2'-MOE 51

552432 2-9-5 2'-MOE 57

552433 2-9-5 2'-MOE 54

552434 2-9-5 2'-MOE 44

552435 2-9-5 2'-MOE 46

552436 2-9-5 2'-MOE 36

552437 2-9-5 2'-MOE 27

552438 2-9-5 2'-MOE 27

552439 2-9-5 2'-MOE 13

Table 32

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000138_0001
552506 4-9-3 2'-MOE 54

552507 4-9-3 2'-MOE 34

552508 4-9-3 2'-MOE 34

552509 4-9-3 2'-MOE 48

552510 4-9-3 2'-MOE 50

552455 3-9-4 2'-MOE 66

55251 1 4-9-3 2'-MOE 66

552456 3-9-4 2'-MOE 64

552512 4-9-3 2'-MOE 62

552457 3-9-4 2'-MOE 14

552513 4-9-3 2'-MOE 56

552458 3-9-4 2'-MOE 59

552514 4-9-3 2'-MOE 52

552459 3-9-4 2'-MOE 69

552515 4-9-3 2'-MOE 57

552460 3-9-4 2'-MOE 0

552516 4-9-3 2'-MOE 54

552461 3-9-4 2'-MOE 20

552517 4-9-3 2'-MOE 52

552462 3-9-4 2'-MOE 46

552518 4-9-3 2'-MOE 34

552463 3-9-4 2'-MOE 48

552519 4-9-3 2'-MOE 44

552464 3-9-4 2'-MOE 81

552520 4-9-3 2'-MOE 69

552465 3-9-4 2'-MOE 84

552521 4-9-3 2'-MOE 80

552466 3-9-4 2'-MOE 75

552522 4-9-3 2'-MOE 76

552467 3-9-4 2'-MOE 65

552523 4-9-3 2'-MOE 71

552468 3-9-4 2'-MOE 53

552524 4-9-3 2'-MOE 43

552469 3-9-4 2'-MOE 51

552525 4-9-3 2'-MOE 57

552470 3-9-4 2'-MOE 46

552526 4-9-3 2'-MOE 60

552471 3-9-4 2'-MOE 54

552527 4-9-3 2'-MOE 72

552472 3-9-4 2'-MOE 78

552528 4-9-3 2'-MOE 78

552473 3-9-4 2'-MOE 67

552529 4-9-3 2'-MOE 77 552474 3-9-4 2'-MOE 79

552530 4-9-3 2'-MOE 78

552475 3-9-4 2'-MOE 74

552531 4-9-3 2'-MOE 68

552476 3-9-4 2'-MOE 52

552477 3-9-4 2'-MOE 76

552478 3-9-4 2'-MOE 70

552479 3-9-4 2'-MOE 67

552480 3-9-4 2'-MOE 68

552481 3-9-4 2'-MOE 57

552482 3-9-4 2'-MOE 51

552483 3-9-4 2'-MOE 48

552484 3-9-4 2'-MOE 58

552485 3-9-4 2'-MOE 51

552486 3-9-4 2'-MOE 55

552487 3-9-4 2'-MOE 62

552488 3-9-4 2'-MOE 51

552489 3-9-4 2'-MOE 49

552490 3-9-4 2'-MOE 51

552491 3-9-4 2'-MOE 51

552492 3-9-4 2'-MOE 38

552493 3-9-4 2'-MOE 52

552494 3-9-4 2'-MOE 17

552495 3-9-4 2'-MOE 49

Table 33

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000140_0001
552564 5-9-2 2'-MOE 41

552565 5-9-2 2'-MOE 54

552566 5-9-2 2'-MOE 56

552567 5-9-2 2'-MOE 71

552568 5-9-2 2'-MOE 64

552569 5-9-2 2'-MOE 59

552570 5-9-2 2'-MOE 60

552571 5-9-2 2'-MOE 55

552572 5-9-2 2'-MOE 60

552573 5-9-2 2'-MOE 24

552574 5-9-2 2'-MOE 34

552575 5-9-2 2'-MOE 36

552576 5-9-2 2'-MOE 67

552577 5-9-2 2'-MOE 64

552578 5-9-2 2'-MOE 75

552579 5-9-2 2'-MOE 75

552580 5-9-2 2'-MOE 59

552581 5-9-2 2'-MOE 54

552582 5-9-2 2'-MOE 61

552583 5-9-2 2'-MOE 69

552584 5-9-2 2'-MOE 74

552585 5-9-2 2'-MOE 62

552586 5-9-2 2'-MOE 79

552587 5-9-2 2'-MOE 71

552532 4-9-3 2'-MOE 48

552588 5-9-2 2'-MOE 70

552533 4-9-3 2'-MOE 43

552589 5-9-2 2'-MOE 59

552534 4-9-3 2'-MOE 62

552590 5-9-2 2'-MOE 70

552535 4-9-3 2'-MOE 55

552591 5-9-2 2'-MOE 51

552536 4-9-3 2'-MOE 3

552592 5-9-2 2'-MOE 50

552537 4-9-3 2'-MOE 14

552593 5-9-2 2'-MOE 46

552538 4-9-3 2'-MOE 52

552594 5-9-2 2'-MOE 55

552539 4-9-3 2'-MOE 47

552595 5-9-2 2'-MOE 60

552540 4-9-3 2'-MOE 60

552596 5-9-2 2'-MOE 63

552541 4-9-3 2'-MOE 60 552597 5-9-2 2'-MOE 61

552542 4-9-3 2'-MOE 64

552598 5-9-2 2'-MOE 57

552543 4-9-3 2'-MOE 46

552600 5-9-2 2'-MOE 59

552544 4-9-3 2'-MOE 53

552602 5-9-2 2'-MOE 6

552545 4-9-3 2'-MOE 33

552604 5-9-2 2'-MOE 47

552546 4-9-3 2'-MOE 42

552606 5-9-2 2'-MOE 53

552547 4-9-3 2'-MOE 51

552608 5-9-2 2'-MOE 53

552548 4-9-3 2'-MOE 52

552610 5-9-2 2'-MOE 47

552549 4-9-3 2'-MOE 38

552612 5-9-2 2'-MOE 39

552550 4-9-3 2'-MOE 19

552614 5-9-2 2'-MOE 24

552551 4-9-3 2'-MOE 24

552616 5-9-2 2'-MOE 15

Table 34

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000142_0001
552046 7- 10-3 2'-MOE 93

552015 6-10-4 2'-MOE 70

552047 7- 10-3 2'-MOE 77

552016 6-10-4 2'-MOE 61

552048 7- 10-3 2'-MOE 66

552017 6-10-4 2'-MOE 73

552049 7- 10-3 2'-MOE 73

552018 6-10-4 2'-MOE 98

552050 7- 10-3 2'-MOE 98

552019 6-10-4 2'-MOE 98

552051 7- 10-3 2'-MOE 99

551986 4-10-6 2'-MOE 92

552020 6-10-4 2'-MOE 97

552052 7- 10-3 2'-MOE 98

551987 4-10-6 2'-MOE 95

552021 6-10-4 2'-MOE 97

552053 7- 10-3 2'-MOE 98

551988 4-10-6 2'-MOE 50

552005 5-10-5 2'-MOE 99

552022 6-10-4 2'-MOE 99

552054 7- 10-3 2'-MOE 99

551989 4-10-6 2'-MOE 96

552023 6-10-4 2'-MOE 99

552055 7- 10-3 2'-MOE 98

551990 4-10-6 2'-MOE 86

552024 6-10-4 2'-MOE 89

552056 7- 10-3 2'-MOE 88

551991 4-10-6 2'-MOE 0

552025 6-10-4 2'-MOE 90

552057 7- 10-3 2'-MOE 92

551992 4-10-6 2'-MOE 72

552026 6-10-4 2'-MOE 88

552058 7- 10-3 2'-MOE 86

551993 4-10-6 2'-MOE 82

552027 6-10-4 2'-MOE 87

552059 7- 10-3 2'-MOE 88

551994 4-10-6 2'-MOE 85

552028 6-10-4 2'-MOE 83

552060 7- 10-3 2'-MOE 82

551995 4-10-6 2'-MOE 84

552029 6-10-4 2'-MOE 88

552061 7- 10-3 2'-MOE 85

551996 4-10-6 2'-MOE 87 552030 6-10-4 2'-MOE 88

552062 7- 10-3 2'-MOE 85

551997 4-10-6 2'-MOE 83

552031 6-10-4 2'-MOE 82

551998 4-10-6 2'-MOE 85

552032 6-10-4 2'-MOE 87

551999 4-10-6 2'-MOE 82

552033 6-10-4 2'-MOE 87

552000 4-10-6 2'-MOE 83

552006 5-10-5 2'-MOE 88

552034 6-10-4 2'-MOE 89

552001 4-10-6 2'-MOE 65

552035 6-10-4 2'-MOE 60

552002 4-10-6 2'-MOE 63

552036 6-10-4 2'-MOE 65

552003 4-10-6 2'-MOE 65

552037 6-10-4 2'-MOE 58

552004 4-10-6 2'-MOE 58

552038 6-10-4 2'-MOE 70

Table 35

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000144_0001
552178 3-9-5 2'-MOE 66

552179 3-9-5 2'-MOE 70

552180 3-9-5 2'-MOE 66

552181 3-9-5 2'-MOE 51

552182 3-9-5 2'-MOE 69

552183 3-9-5 2'-MOE 69

552184 3-9-5 2'-MOE 43

552185 3-9-5 2'-MOE 66

552186 3-9-5 2'-MOE 54

552187 3-9-5 2'-MOE 74

552188 3-9-5 2'-MOE 78

552189 3-9-5 2'-MOE 57

552190 3-9-5 2'-MOE 39

552191 3-9-5 2'-MOE 60

552192 3-9-5 2'-MOE 85

552193 3-9-5 2'-MOE 86

552194 3-9-5 2'-MOE 68

552195 3-9-5 2'-MOE 73

552196 3-9-5 2'-MOE 60

552197 3-9-5 2'-MOE 60

552198 3-9-5 2'-MOE 61

552199 3-9-5 2'-MOE 89

552200 3-9-5 2'-MOE 85

552201 3-9-5 2'-MOE 81

552202 3-9-5 2'-MOE 76

552203 3-9-5 2'-MOE 74

552204 3-9-5 2'-MOE 71

552151 2-9-6 2'-MOE 77

552205 3-9-5 2'-MOE 78

552152 2-9-6 2'-MOE 72

552206 3-9-5 2'-MOE 77

552153 2-9-6 2'-MOE 67

552207 3-9-5 2'-MOE 81

552154 2-9-6 2'-MOE 56

552208 3-9-5 2'-MOE 70

552155 2-9-6 2'-MOE 61

552209 3-9-5 2'-MOE 63

552156 2-9-6 2'-MOE 20

552210 3-9-5 2'-MOE 75

552157 2-9-6 2'-MOE 39

55221 1 3-9-5 2'-MOE 75

552158 2-9-6 2'-MOE 70

552212 3-9-5 2'-MOE 67 552159 2-9-6 2'-MOE 74

552213 3-9-5 2'-MOE 70

552160 2-9-6 2'-MOE 78

552214 3-9-5 2'-MOE 79

552161 2-9-6 2'-MOE 56

552215 3-9-5 2'-MOE 61

552162 2-9-6 2'-MOE 64

552216 3-9-5 2'-MOE 62

552163 2-9-6 2'-MOE 71

552217 3-9-5 2'-MOE 58

552164 2-9-6 2'-MOE 52

552218 3-9-5 2'-MOE 56

552165 2-9-6 2'-MOE 53

552219 3-9-5 2'-MOE 33

552166 2-9-6 2'-MOE 41

552220 3-9-5 2'-MOE 53

552167 2-9-6 2'-MOE 54

552221 3-9-5 2'-MOE 31

Table 36

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000146_0001
552125 2-9-6 2'-MOE 73

552077 8-10-2 2'-MOE 83

552126 2-9-6 2'-MOE 64

552078 8-10-2 2'-MOE 80

552127 2-9-6 2'-MOE 72

552079 8-10-2 2'-MOE 86

552128 2-9-6 2'-MOE 76

552080 8-10-2 2'-MOE 83

552129 2-9-6 2'-MOE 72

552131 2-9-6 2'-MOE 61

552132 2-9-6 2'-MOE 73

552133 2-9-6 2'-MOE 75

552081 8-10-2 2'-MOE 76

552134 2-9-6 2'-MOE 58

552135 2-9-6 2'-MOE 67

552136 2-9-6 2'-MOE 65

552137 2-9-6 2'-MOE 55

552082 8-10-2 2'-MOE 98

552138 2-9-6 2'-MOE 82

552083 8-10-2 2'-MOE 99

552139 2-9-6 2'-MOE 86

552084 8-10-2 2'-MOE 99

552140 2-9-6 2'-MOE 74

552085 8-10-2 2'-MOE 100

552141 2-9-6 2'-MOE 67

552086 8-10-2 2'-MOE 100

552142 2-9-6 2'-MOE 45

552087 8-10-2 2'-MOE 100

552143 2-9-6 2'-MOE 68

552144 2-9-6 2'-MOE 78

552145 2-9-6 2'-MOE 88

552146 2-9-6 2'-MOE 81

552088 8-10-2 2'-MOE 95

552147 2-9-6 2'-MOE 88

552089 8-10-2 2'-MOE 93

552148 2-9-6 2'-MOE 79

552090 8-10-2 2'-MOE 87

552149 2-9-6 2'-MOE 81

552091 8-10-2 2'-MOE 88

552092 8-10-2 2'-MOE 90

552093 8-10-2 2'-MOE 91

552094 8-10-2 2'-MOE 88

552063 7-10-3 2'-MOE 81 552095 8-10-2 2'-MOE 89

552064 7-10-3 2'-MOE 85

552096 8-10-2 2'-MOE 92

552065 7-10-3 2'-MOE 86

552097 8-10-2 2'-MOE 93

552066 7-10-3 2'-MOE 33

552098 8-10-2 2'-MOE 88

552067 7-10-3 2'-MOE 50

552099 8-10-2 2'-MOE 70

552068 7-10-3 2'-MOE 73

552100 8-10-2 2'-MOE 70

552069 7-10-3 2'-MOE 73

552101 8-10-2 2'-MOE 76

552070 7-10-3 2'-MOE 71

552102 8-10-2 2'-MOE 64

Table 37

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000148_0001
552351 6-9-2 2'-MOE 60

552352 6-9-2 2'-MOE 53

552353 6-9-2 2'-MOE 0

552354 6-9-2 2'-MOE 83

552355 6-9-2 2'-MOE 90

552356 6-9-2 2'-MOE 0

552357 6-9-2 2'-MOE 45

552358 6-9-2 2'-MOE 74

552359 6-9-2 2'-MOE 72

552360 6-9-2 2'-MOE 87

552361 6-9-2 2'-MOE 96

552308 5-9-3 2'-MOE 81

552362 6-9-2 2'-MOE 92

552309 5-9-3 2'-MOE 77

552363 6-9-2 2'-MOE 92

552310 5-9-3 2'-MOE 80

552364 6-9-2 2'-MOE 87

55231 1 5-9-3 2'-MOE 13

552365 6-9-2 2'-MOE 84

552150 2-9-6 2'-MOE 73

552312 5-9-3 2'-MOE 77

552366 6-9-2 2'-MOE 87

552313 5-9-3 2'-MOE 64

552367 6-9-2 2'-MOE 85

552314 5-9-3 2'-MOE 73

552368 6-9-2 2'-MOE 77

552315 5-9-3 2'-MOE 75

552369 6-9-2 2'-MOE 75

552316 5-9-3 2'-MOE 64

552370 6-9-2 2'-MOE 63

552317 5-9-3 2'-MOE 99

552371 6-9-2 2'-MOE 81

552318 5-9-3 2'-MOE 76

552372 6-9-2 2'-MOE 65

552319 5-9-3 2'-MOE 55

552373 6-9-2 2'-MOE 74

552320 5-9-3 2'-MOE 68

552374 6-9-2 2'-MOE 78

552321 5-9-3 2'-MOE 74

552375 6-9-2 2'-MOE 81

552322 5-9-3 2'-MOE 73

552376 6-9-2 2'-MOE 78

552323 5-9-3 2'-MOE 75 552377 6-9-2 2'-MOE 70

552324 5-9-3 2'-MOE 0

552378 6-9-2 2'-MOE 72

552325 5-9-3 2'-MOE 70

552379 6-9-2 2'-MOE 74

552326 5-9-3 2'-MOE 63

552380 6-9-2 2'-MOE 53

552327 5-9-3 2'-MOE 30

552381 6-9-2 2'-MOE 26

552328 5-9-3 2'-MOE 25

552382 6-9-2 2'-MOE 13

552329 5-9-3 2'-MOE 33

552383 6-9-2 2'-MOE 5

Table 38

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3370

Figure imgf000150_0001
551917 2-10-8 2'-MOE 58

551949 3-10-7 2'-MOE 64

551981 4-10-6 2'-MOE 63

551918 2-10-8 2'-MOE 59

551950 3-10-7 2'-MOE 71

551982 4-10-6 2'-MOE 63

551919 2-10-8 2'-MOE 76

551951 3-10-7 2'-MOE 71

551983 4-10-6 2'-MOE 73

551920 2-10-8 2'-MOE 68

551952 3-10-7 2'-MOE 76

551984 4-10-6 2'-MOE 81

551921 2-10-8 2'-MOE 83

551953 3-10-7 2'-MOE 82

551985 4-10-6 2'-MOE 76

551922 2-10-8 2'-MOE 73

551954 3-10-7 2'-MOE 68

551923 2-10-8 2'-MOE 59

551955 3-10-7 2'-MOE 71

551924 2-10-8 2'-MOE 80

551956 3-10-7 2'-MOE 80

551925 2-10-8 2'-MOE 82

551957 3-10-7 2'-MOE 88

551926 2-10-8 2'-MOE 71

551958 3-10-7 2'-MOE 74

551927 2-10-8 2'-MOE 68

551959 3-10-7 2'-MOE 69

551928 2-10-8 2'-MOE 69

551960 3-10-7 2'-MOE 62

551929 2-10-8 2'-MOE 54

551961 3-10-7 2'-MOE 20

551930 2-10-8 2'-MOE 53

551962 3-10-7 2'-MOE 60

551931 2-10-8 2'-MOE 47

551963 3-10-7 2'-MOE 63

551932 2-10-8 2'-MOE 68

551964 3-10-7 2'-MOE 56

551933 2-10-8 2'-MOE 72

551965 3-10-7 2'-MOE 67

551934 2-10-8 2'-MOE 64

551966 3-10-7 2'-MOE 73

551935 2-10-8 2'-MOE 68

551967 3-10-7 2'-MOE 60 551936 2-10-8 2'-MOE 67

551968 3-10-7 2'-MOE 63

551937 2-10-8 2'-MOE 47

551969 3-10-7 2'-MOE 36

551938 2-10-8 2'-MOE 41

551970 3-10-7 2'-MOE 43

551939 2-10-8 2'-MOE 53

551971 3-10-7 2'-MOE 55

551940 2-10-8 2'-MOE 50

551972 3-10-7 2'-MOE 58

Table 39

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000152_0001
551918 2-10-8 2'-MOE 53

551950 3-10-7 2'-MOE 57

551982 4-10-6 2'-MOE 57

551919 2-10-8 2'-MOE 65

551951 3-10-7 2'-MOE 57

551983 4-10-6 2'-MOE 53

551920 2-10-8 2'-MOE 57

551952 3-10-7 2'-MOE 67

551984 4-10-6 2'-MOE 62

551921 2-10-8 2'-MOE 60

551953 3-10-7 2'-MOE 57

551985 4-10-6 2'-MOE 58

551922 2-10-8 2'-MOE 63

551954 3-10-7 2'-MOE 61

551923 2-10-8 2'-MOE 50

551955 3-10-7 2'-MOE 44

551924 2-10-8 2'-MOE 52

551956 3-10-7 2'-MOE 46

551925 2-10-8 2'-MOE 54

551957 3-10-7 2'-MOE 51

551926 2-10-8 2'-MOE 70

551958 3-10-7 2'-MOE 72

551927 2-10-8 2'-MOE 60

551959 3-10-7 2'-MOE 61

551928 2-10-8 2'-MOE 57

551960 3-10-7 2'-MOE 58

551929 2-10-8 2'-MOE 49

551961 3-10-7 2'-MOE 26

551930 2-10-8 2'-MOE 54

551962 3-10-7 2'-MOE 57

551931 2-10-8 2'-MOE 46

551963 3-10-7 2'-MOE 56

551932 2-10-8 2'-MOE 57

551964 3-10-7 2'-MOE 53

551933 2-10-8 2'-MOE 65

551965 3-10-7 2'-MOE 54

551934 2-10-8 2'-MOE 58

551966 3-10-7 2'-MOE 69

551935 2-10-8 2'-MOE 63

551967 3-10-7 2'-MOE 53

551936 2-10-8 2'-MOE 67

551968 3-10-7 2'-MOE 60

551937 2-10-8 2'-MOE 51 551969 3-10-7 2'-MOE 42

551938 2-10-8 2'-MOE 40

551970 3-10-7 2'-MOE 38

551939 2-10-8 2'-MOE 32

551971 3-10-7 2'-MOE 46

551940 2-10-8 2'-MOE 39

551972 3-10-7 2'-MOE 51

Table 40

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000154_0001
552241 4-9-4 2'-MOE 47

552295 5-9-3 2'-MOE 63

552242 4-9-4 2'-MOE 61

552296 5-9-3 2'-MOE 61

552243 4-9-4 2'-MOE 55

552297 5-9-3 2'-MOE 52

552244 4-9-4 2'-MOE 45

552298 5-9-3 2'-MOE 27

552245 4-9-4 2'-MOE 41

552299 5-9-3 2'-MOE 32

552246 4-9-4 2'-MOE 67

552300 5-9-3 2'-MOE 57

552247 4-9-4 2'-MOE 74

552301 5-9-3 2'-MOE 76

552248 4-9-4 2'-MOE 65

552302 5-9-3 2'-MOE 68

552249 4-9-4 2'-MOE 38

552303 5-9-3 2'-MOE 59

552250 4-9-4 2'-MOE 43

552304 5-9-3 2'-MOE 30

552251 4-9-4 2'-MOE 52

552305 5-9-3 2'-MOE 49

552252 4-9-4 2'-MOE 51

552306 5-9-3 2'-MOE 56

552253 4-9-4 2'-MOE 47

552307 5-9-3 2'-MOE 49

552254 4-9-4 2'-MOE 50

552255 4-9-4 2'-MOE 64

552256 4-9-4 2'-MOE 57

552257 4-9-4 2'-MOE 51

552258 4-9-4 2'-MOE 62

552259 4-9-4 2'-MOE 59

552260 4-9-4 2'-MOE 56

552261 4-9-4 2'-MOE 54

552262 4-9-4 2'-MOE 47

552263 4-9-4 2'-MOE 45

552264 4-9-4 2'-MOE 52

552265 4-9-4 2'-MOE 58

552266 4-9-4 2'-MOE 54

552267 4-9-4 2'-MOE 43

552268 4-9-4 2'-MOE 57

552269 4-9-4 2'-MOE 34

552270 4-9-4 2'-MOE 37

54 552271 4-9-4 2'-MOE 42

552272 4-9-4 2'-MOE 36

552273 4-9-4 2'-MOE 25

552274 4-9-4 2'-MOE 11

552275 4-9-4 2'-MOE 38

Table 41

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000156_0001
552346 6-9-2 2'-MOE 0

552397 2-9-5 2'-MOE 37

552453 3-9-4 2'-MOE 45

552398 2-9-5 2'-MOE 42

552454 3-9-4 2'-MOE 39

552399 2-9-5 2'-MOE 34

552400 2-9-5 2'-MOE 47

552401 2-9-5 2'-MOE 53

552402 2-9-5 2'-MOE 47

552403 2-9-5 2'-MOE 70

552404 2-9-5 2'-MOE 44

552405 2-9-5 2'-MOE 0

552406 2-9-5 2'-MOE 25

552407 2-9-5 2'-MOE 23

552408 2-9-5 2'-MOE 73

552409 2-9-5 2'-MOE 71

552410 2-9-5 2'-MOE 52

55241 1 2-9-5 2'-MOE 62

552412 2-9-5 2'-MOE 50

552413 2-9-5 2'-MOE 55

552414 2-9-5 2'-MOE 64

552415 2-9-5 2'-MOE 45

552416 2-9-5 2'-MOE 45

552417 2-9-5 2'-MOE 37

552418 2-9-5 2'-MOE 73

552419 2-9-5 2'-MOE 68

552420 2-9-5 2'-MOE 64

552421 2-9-5 2'-MOE 54

552422 2-9-5 2'-MOE 60

552423 2-9-5 2'-MOE 62

552424 2-9-5 2'-MOE 60

552425 2-9-5 2'-MOE 46

552426 2-9-5 2'-MOE 48

552427 2-9-5 2'-MOE 36

552428 2-9-5 2'-MOE 57

552429 2-9-5 2'-MOE 36

552430 2-9-5 2'-MOE 42

552431 2-9-5 2'-MOE 60

552432 2-9-5 2'-MOE 44

552433 2-9-5 2'-MOE 55

552434 2-9-5 2'-MOE 46

552435 2-9-5 2'-MOE 47

552436 2-9-5 2'-MOE 25

56 552437 2-9-5 2'-MOE 19

552438 2-9-5 2'-MOE 25

552439 2-9-5 2'-MOE 22

Table 42

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000158_0001
552520 4-9-3 2'-MOE 61

552465 3-9-4 2'-MOE 68

552521 4-9-3 2'-MOE 68

552466 3-9-4 2'-MOE 76

552522 4-9-3 2'-MOE 71

552467 3-9-4 2'-MOE 72

552523 4-9-3 2'-MOE 73

552468 3-9-4 2'-MOE 50

552524 4-9-3 2'-MOE 49

552469 3-9-4 2'-MOE 65

552525 4-9-3 2'-MOE 45

552470 3-9-4 2'-MOE 58

552526 4-9-3 2'-MOE 39

552471 3-9-4 2'-MOE 30

552527 4-9-3 2'-MOE 39

552472 3-9-4 2'-MOE 43

552528 4-9-3 2'-MOE 43

552473 3-9-4 2'-MOE 25

552529 4-9-3 2'-MOE 50

552474 3-9-4 2'-MOE 70

552530 4-9-3 2'-MOE 73

552475 3-9-4 2'-MOE 64

552531 4-9-3 2'-MOE 62

552476 3-9-4 2'-MOE 50

552477 3-9-4 2'-MOE 66

552478 3-9-4 2'-MOE 68

552479 3-9-4 2'-MOE 60

552480 3-9-4 2'-MOE 58

552481 3-9-4 2'-MOE 54

552482 3-9-4 2'-MOE 44

552483 3-9-4 2'-MOE 17

552484 3-9-4 2'-MOE 64

552485 3-9-4 2'-MOE 56

552486 3-9-4 2'-MOE 26

552487 3-9-4 2'-MOE 42

552488 3-9-4 2'-MOE 35

552489 3-9-4 2'-MOE 46

552490 3-9-4 2'-MOE 41

552491 3-9-4 2'-MOE 38

552492 3-9-4 2'-MOE 47

552493 3-9-4 2'-MOE 49

552494 3-9-4 2'-MOE 22

552495 3-9-4 2'-MOE 0 Table 43

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000160_0001
552588 5-9-2 2'-MOE 67

552533 4-9-3 2'-MOE 52

552589 5-9-2 2'-MOE 47

552534 4-9-3 2'-MOE 71

552590 5-9-2 2'-MOE 58

552535 4-9-3 2'-MOE 59

552591 5-9-2 2'-MOE 46

552536 4-9-3 2'-MOE 19

552592 5-9-2 2'-MOE 44

552537 4-9-3 2'-MOE 26

552593 5-9-2 2'-MOE 39

552538 4-9-3 2'-MOE 54

552594 5-9-2 2'-MOE 52

552539 4-9-3 2'-MOE 50

552595 5-9-2 2'-MOE 57

552540 4-9-3 2'-MOE 60

552596 5-9-2 2'-MOE 58

552541 4-9-3 2'-MOE 68

552597 5-9-2 2'-MOE 52

552542 4-9-3 2'-MOE 63

552598 5-9-2 2'-MOE 51

552543 4-9-3 2'-MOE 44

552600 5-9-2 2'-MOE 51

552544 4-9-3 2'-MOE 45

552602 5-9-2 2'-MOE 13

552545 4-9-3 2'-MOE 42

552604 5-9-2 2'-MOE 42

552546 4-9-3 2'-MOE 46

552606 5-9-2 2'-MOE 42

552547 4-9-3 2'-MOE 38

552608 5-9-2 2'-MOE 37

552548 4-9-3 2'-MOE 49

552610 5-9-2 2'-MOE 41

552549 4-9-3 2'-MOE 34

552612 5-9-2 2'-MOE 23

552550 4-9-3 2'-MOE 13

552614 5-9-2 2'-MOE 1 1

552551 4-9-3 2'-MOE 8

552616 5-9-2 2'-MOE 6 Table 44

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000162_0001
552023 6-10-4 2'-MOE 78

552055 7-10-3 2'-MOE 57

551990 4-10-6 2'-MOE 83

552024 6-10-4 2'-MOE 89

552056 7-10-3 2'-MOE 82

551991 4-10-6 2'-MOE 0

552025 6-10-4 2'-MOE 89

552057 7-10-3 2'-MOE 89

551992 4-10-6 2'-MOE 67

552026 6-10-4 2'-MOE 84

552058 7-10-3 2'-MOE 82

551993 4-10-6 2'-MOE 78

552027 6-10-4 2'-MOE 85

552059 7-10-3 2'-MOE 85

551994 4-10-6 2'-MOE 82

552028 6-10-4 2'-MOE 82

552060 7-10-3 2'-MOE 74

551995 4-10-6 2'-MOE 81

552029 6-10-4 2'-MOE 81

552061 7-10-3 2'-MOE 81

551996 4-10-6 2'-MOE 79

552030 6-10-4 2'-MOE 86

552062 7-10-3 2'-MOE 85

551997 4-10-6 2'-MOE 80

552031 6-10-4 2'-MOE 86

551998 4-10-6 2'-MOE 74

552032 6-10-4 2'-MOE 78

551999 4-10-6 2'-MOE 79

552033 6-10-4 2'-MOE 80

552000 4-10-6 2'-MOE 84

552006 5-10-5 2'-MOE 86

552034 6-10-4 2'-MOE 81

552001 4-10-6 2'-MOE 66

552035 6-10-4 2'-MOE 55

552002 4-10-6 2'-MOE 54

552036 6-10-4 2'-MOE 58

552003 4-10-6 2'-MOE 50

552037 6-10-4 2'-MOE 43

552004 4-10-6 2'-MOE 56

552038 6-10-4 2'-MOE 66 Table 45

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000164_0001
552199 3-9-5 2'-MOE 57

552200 3-9-5 2'-MOE 52

552201 3-9-5 2'-MOE 73

552202 3-9-5 2'-MOE 60

552203 3-9-5 2'-MOE 60

552204 3-9-5 2'-MOE 63

552151 2-9-6 2'-MOE 71

552205 3-9-5 2'-MOE 64

552152 2-9-6 2'-MOE 69

552206 3-9-5 2'-MOE 71

552153 2-9-6 2'-MOE 63

552207 3-9-5 2'-MOE 71

552154 2-9-6 2'-MOE 56

552208 3-9-5 2'-MOE 52

552155 2-9-6 2'-MOE 61

552209 3-9-5 2'-MOE 50

552156 2-9-6 2'-MOE 40

552210 3-9-5 2'-MOE 66

552157 2-9-6 2'-MOE 45

55221 1 3-9-5 2'-MOE 63

552158 2-9-6 2'-MOE 66

552212 3-9-5 2'-MOE 62

552159 2-9-6 2'-MOE 68

552213 3-9-5 2'-MOE 64

552160 2-9-6 2'-MOE 78

552214 3-9-5 2'-MOE 72

552161 2-9-6 2'-MOE 57

552215 3-9-5 2'-MOE 54

552162 2-9-6 2'-MOE 54

552216 3-9-5 2'-MOE 49

552163 2-9-6 2'-MOE 65

552217 3-9-5 2'-MOE 50

552164 2-9-6 2'-MOE 48

552218 3-9-5 2'-MOE 39

552165 2-9-6 2'-MOE 46

552219 3-9-5 2'-MOE 41

552166 2-9-6 2'-MOE 42

552220 3-9-5 2'-MOE 32

552167 2-9-6 2'-MOE 47

552221 3-9-5 2'-MOE 33 Table 46

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000166_0001
552138 2-9-6 2'-MOE 80

552083 8-10-2 2'-MOE 85

552139 2-9-6 2'-MOE 80

552084 8-10-2 2'-MOE 86

552140 2-9-6 2'-MOE 70

552085 8-10-2 2'-MOE 83

552141 2-9-6 2'-MOE 72

552086 8-10-2 2'-MOE 83

552142 2-9-6 2'-MOE 58

552087 8-10-2 2'-MOE 77

552143 2-9-6 2'-MOE 70

552144 2-9-6 2'-MOE 66

552145 2-9-6 2'-MOE 78

552146 2-9-6 2'-MOE 63

552088 8-10-2 2'-MOE 90

552147 2-9-6 2'-MOE 80

552089 8-10-2 2'-MOE 87

552148 2-9-6 2'-MOE 74

552090 8-10-2 2'-MOE 85

552149 2-9-6 2'-MOE 79

552091 8-10-2 2'-MOE 84

552092 8-10-2 2'-MOE 86

552093 8-10-2 2'-MOE 82

552094 8-10-2 2'-MOE 84

552063 7-10-3 2'-MOE 79

552095 8-10-2 2'-MOE 85

552064 7-10-3 2'-MOE 83

552096 8-10-2 2'-MOE 88

552065 7-10-3 2'-MOE 86

552097 8-10-2 2'-MOE 90

552066 7-10-3 2'-MOE 35

552098 8-10-2 2'-MOE 86

552067 7-10-3 2'-MOE 53

552099 8-10-2 2'-MOE 66

552068 7-10-3 2'-MOE 70

552100 8-10-2 2'-MOE 67

552069 7-10-3 2'-MOE 68

552101 8-10-2 2'-MOE 65

552070 7-10-3 2'-MOE 64

552102 8-10-2 2'-MOE 54 Table 47

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotidesmeasured with RTS3371

Figure imgf000168_0001
552364 6-9-2 2'-MOE 66

55231 1 5-9-3 2'-MOE 38

552365 6-9-2 2'-MOE 64

552150 2-9-6 2'-MOE 68

552312 5-9-3 2'-MOE 75

552366 6-9-2 2'-MOE 55

552313 5-9-3 2'-MOE 66

552367 6-9-2 2'-MOE 67

552314 5-9-3 2'-MOE 56

552368 6-9-2 2'-MOE 41

552315 5-9-3 2'-MOE 46

552369 6-9-2 2'-MOE 52

552316 5-9-3 2'-MOE 55

552370 6-9-2 2'-MOE 35

552317 5-9-3 2'-MOE 53

552371 6-9-2 2'-MOE 58

552318 5-9-3 2'-MOE 59

552372 6-9-2 2'-MOE 68

552319 5-9-3 2'-MOE 56

552373 6-9-2 2'-MOE 63

552320 5-9-3 2'-MOE 62

552374 6-9-2 2'-MOE 70

552321 5-9-3 2'-MOE 63

552375 6-9-2 2'-MOE 64

552322 5-9-3 2'-MOE 52

552376 6-9-2 2'-MOE 58

552323 5-9-3 2'-MOE 45

552377 6-9-2 2'-MOE 42

552324 5-9-3 2'-MOE 49

552378 6-9-2 2'-MOE 37

552325 5-9-3 2'-MOE 48

552379 6-9-2 2'-MOE 57

552326 5-9-3 2'-MOE 50

552380 6-9-2 2'-MOE 48

552327 5-9-3 2'-MOE 13

552381 6-9-2 2'-MOE 22

552328 5-9-3 2'-MOE 9

552382 6-9-2 2'-MOE 20

552329 5-9-3 2'-MOE 18

552383 6-9-2 2'-MOE 18 Example 10: Dose-dependent antisense inhibition of Target-Z mRNA in HepG2 cells

Antisense oligonucleotides from the study described in Example 52 exhibiting in vitro inhibition of Target-Z mRNA were selected and tested at various doses in HepG2 cells. Cells were plated at a density of 28,000 cells per well and transfected using LipofectAMINE2000® with 9.26 nM, 27.78 nM, 83.33 nM, and 250.00 nM concentrations of antisense oligonucleotide, as specified in Table 48. After a treatment period of approximately 16 hours, RNA was isolated from the cells and Target-Z mRNA levels were measured by quantitative real-time PCR. Target-Z primer probe set RTS3371 was used to measure mRNA levels. Target- Z mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target-Z, relative to untreated control cells.

As illustrated in Table 48, Target-Z mRNA levels were reduced in a dose-dependent manner in antisense oligonucleotide treated cells, 'n/a' indicates that the data for that dosage is not available.

Table 48

Dose-dependent antisense inhibition of human Target-Z in HepG2 cells

Figure imgf000170_0001
552022 6 35 57 79

552023 1 1 33 59 81

552024 15 43 69 91

552025 17 35 69 87

552026 14 26 66 86

552027 3 46 62 88

552028 9 43 58 78

552029 8 40 72 89

552030 18 48 77 92

552031 0 38 66 89

552032 42 48 80 88

552033 2 40 64 84

552034 6 40 70 81

552039 2 33 56 83

552044 19 30 63 84

552046 4 21 47 77

552050 15 44 70 92

552051 8 33 69 90

552052 17 38 71 91

552053 0 40 59 86

552054 7 15 58 75

552056 19 62 86 92

552057 1 1 33 69 86

552058 30 55 79 90

552059 1 1 25 69 90

552060 9 32 61 86

552061 6 40 69 88

552062 22 48 75 89

552064 23 49 69 90

552065 10 8 69 86

552069 1 1 4 28 60

552073 9 31 62 78

552075 21 18 33 65

552077 0 17 40 72

552079 1 12 44 70

552080 3 12 34 69

552082 13 29 66 87

552083 24 54 69 88

552084 10 25 48 82

552085 28 35 64 85

552086 0 24 65 84

552088 33 53 77 93

552089 0 41 69 92 552090 17 35 70 87

552091 13 31 69 89

552092 6 23 66 89

552093 0 17 61 89

552094 12 38 65 88

552095 20 42 73 88

552096 n/a 39 66 91

552097 24 43 67 88

552098 0 24 56 85

552101 3 13 28 61

552147 1 1 27 58 80

552160 20 25 69 89

552163 0 21 22 53

552176 16 1 1 40 66

552192 7 38 78 89

552222 0 24 65 79

552247 0 38 69 86

552255 5 27 69 81

552301 5 38 65 86

552309 8 26 62 85

552312 0 4 32 62

552347 2 15 38 75

552348 12 40 42 65

552354 10 35 44 76

552361 2 25 55 74

552363 20 36 54 76

552374 7 4 38 76

552379 0 12 24 46

552403 8 27 54 76

552408 2 25 44 77

552409 6 31 56 80

552418 0 30 72 84

552420 9 34 53 81

552442 4 23 46 56

552466 0 23 56 79

552474 1 1 34 66 87

552477 1 1 22 44 64

552530 25 37 73 87

552559 9 13 29 51

Example 11: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Target-Z transgenic mice were treated with ISIS antisense oligonucleotides selected from studies described above and evaluated for their efficacy in this model.

Treatment

Groups of 12 mice each were injected subcutaneously twice a week for 4 weeks with 50 mg/kg of ISIS 510106, ISIS 5101 16, ISIS 505347, or ISIS 509934. A control group of 12 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and livers were harvested for further analysis.

DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe sets RTS3370, RTS3371, and RTS3372. The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe sets RTS3370 and RTS3371 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. The data is presented in Table 49, expressed as percent inhibition compared to the control group. As shown in Table 49, most of the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control. Results are presented as percent inhibition of Target-Z mRNA or DNA, relative to control.

Table 49

Percent inhibition of Target-Z RNA and DNA in the liver of transgenic mice

Figure imgf000173_0001

Example 12: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Target-Z transgenic mice were treated with ISIS antisense oligonucleotides selected from studies described above and evaluated for their efficacy in this model.

Treatment

Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 50 mg/kg of ISIS 146779, ISIS 505358, ISIS 146786, ISIS 509974, ISIS 509958, or ISIS 509959. A control group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and livers were harvested for further analysis. DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe sets RTS3370. The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe sets RTS3370 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. The data is presented in Table 50, expressed as percent inhibition compared to the control group. As shown in Table 50, most of the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control. Results are presented as percent inhibition of Target-Z mRNA or DNA, relative to control.

Table 50

Percent inhibition of Target-Z RNA and DNA in the liver of transgenic mice

Figure imgf000174_0001

Example 13: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Transgenic mice were treated with ISIS antisense oligonucleotides selected from studies described above and evaluated for their efficacy in this model.

Treatment

Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786, ISIS 552176, and ISIS 552073. One group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe set RTS3371. The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe set RTS3371 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. The data is presented in Table 51. Serum DNA samples were analyzed after the study period. The data is presented in Table 52, expressed relative to the levels measured in the control group. As shown in Tables 51 and 52, the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control. Results are presented as percent inhibition of Target-Z mRNA or DNA, relative to control.

Table 51

Percent inhibition of Target-Z RNA and DNA in transgenic mice

Figure imgf000175_0001

Table 52

Serum levels of Target-Z DNA in transgenic mice, relative to control levels

Figure imgf000175_0002

Liver function

To evaluate the effect of ISIS oligonucleotides on hepatic function, plasma concentrations of ALT were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, NY) (Nyblom, H. et al., Alcohol & Alcoholism 39: 336-339, 2004; Tietz NW (Ed): Clinical Guide to Laboratory Tests, 3rd ed. W. B. Saunders, Philadelphia, PA, 1995). The results are presented in Table 53 expressed in IU/L. Both the ISIS oligonucleotides were considered tolerable in the mice, as demonstrated by their liver transaminase profile.

Table 53

ALT levels (IU/L) of transgenic mice

Figure imgf000175_0003

Example 14: Efficacy of antisense oligonucleotides targeting Target-Z in transgenic mice

Transgenic mice were treated with ISIS antisense oligonucleotides selected from studies described above and evaluated for their efficacy in this model.

Treatment

Groups of 6 mice each were injected subcutaneously twice a week for 4 weeks with 25 mg/kg of ISIS 146786, ISIS 552056, ISIS 552088, and ISIS 552309. One group of 10 mice was injected subcutaneously twice a week for 4 weeks with PBS. Mice were euthanized 48 hours after the last dose, and organs and plasma were harvested for further analysis.

DNA and RNA Analysis

RNA was extracted from liver tissue for real-time PCR analysis of Target-Z DNA, using primer probe set RTS3371. The DNA levels were normalized to picogreen. Target-Z RNA samples were also assayed with primer probe set RTS3371 after RT-PCR analysis. The mRNA levels were normalized to RIBOGREEN®. As shown in Table 54, the antisense oligonucleotides achieved reduction of Target-Z DNA and RNA over the PBS control. Results are presented as percent inhibition of Target-Z mRNA or DNA, relative to control.

Table 54

Percent inhibition of Target-Z DNA and RNA in transgenic mice

Figure imgf000176_0001

Example 15: Antisense inhibition of Target-Z viral mRNA in HepG2 cells by deoxy, MOE and (S)-cEt gapmers

Additional antisense oligonucleotides were designed targeting a Target-Z viral nucleic acid and were tested for their effects on Target-Z mRNA in vitro. Cultured HepG2 cells at a density of 28,000 cells per well were transfected using LipofectAMINE2000® with 100 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Target-Z mRNA levels were measured by quantitative real-time PCR. Viral primer probe sets RTS3370 and RTS3371 and were used to separately measure mRNA levels. Target-Z mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target-Z, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in Table below were designed as MOE gapmers or deoxy, MOE and (S)-cEt gapmers. The 5-10-5 MOE gapmers are 20 nucleosides in length, wherein the central gap segment comprises of ten 2'-deoxynucleosides and is flanked on both sides (in the 5' and 3 ' directions) by wings comprising five nucleosides each. The deoxy, MOE and (S)-cEt gapmers are 16 nucleosides in length wherein the nucleoside have either a MOE sugar modification, an (S)-cEt sugar modification, or a deoxy modification. The 'Chemistry' column describes the sugar modifications of each oligonucleotide, 'k' indicates an (S)-cEt sugar modification; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate (P=S) linkages. All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

Each gapmer listed in Table 55 is targeted to the viral Target-Z genomic sequence.

Table 55

Inhibition of viral Target-Z mRNA levels by chimeric antisense oligonucleotides measured with RTS3370 or

RTS3371

Figure imgf000177_0001
9591 Uniform deoxy 30 14

Example 16: Antisense inhibition of Target-Z viral mRNA in HepG2 cells by deoxy, MOE and (S)-cEt gapmers

Additional antisense oligonucleotides were designed targeting a Target-Z viral nucleic acid and were tested for their effects on Target-Z mRNA in vitro. ISIS 577121, ISIS 577122, ISIS 577123, ISIS 577132, ISIS 577133, and ISIS 577134, disclosed in the study described above, were also included in the assay. Cultured HepG2 cells at a density of 28,000 cells per well were transfected using Cytofectin with 9.375 nM, 18.75 nM, 37.50 nM, 75.00 nM, 150.00 nM, or 300.00 nM antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and Target-Z mRNA levels were measured by quantitative real-time PCR. Viral primer probe set RTS3371 was used to measure mRNA levels. Target-Z mRNA levels were adjusted according to total RNA content, as measured by RIBOGREEN®. Results are presented as percent inhibition of Target-Z, relative to untreated control cells.

The newly designed chimeric antisense oligonucleotides in Tables below were designed as deoxy, MOE and (S)-cEt gapmers. The deoxy, MOE and (S)-cEt gapmers are 16, 17, or 18 nucleosides in length wherein the nucleosides have either a MOE sugar modification, an (S)-cEt sugar modification, or a deoxy modification. The 'Chemistry' column describes the sugar modifications of each oligonucleotide, 'k' indicates an (S)-cEt sugar modification; the number indicates the number of deoxynucleosides; otherwise, 'd' indicates a deoxynucleoside; and 'e' indicates a MOE modification. The internucleoside linkages throughout each gapmer are phosphorothioate (P=S) linkages. All cytosine residues throughout each oligonucleotide are 5-methylcytosines.

Each gapmer listed in Table 56 is targeted to the viral genomic sequence.

Table 56

Chimeric antisense oligonucleotides targeting viral Target-Z genomic sequence

Figure imgf000178_0001
585168 eeekk-7-kkeee

585169 eeek-9-keee

Table 57

Dose dependent inhibition of Target-Z mRNA levels by chimeric antisense oligonucleotides

Figure imgf000179_0001

Table 58

Dose dependent inhibition of Target-Z mRNA levels by chimeric antisense oligonucleotides

Figure imgf000179_0002

Claims

We claim:
1. A compound comprising:
a modified oligonucleotide consisting of 10 to 20 linked nucleosides, wherein the modified oligonucleotide comprises:
a 5 '-wing consisting of 2 to 5 linked nucleosides;
a 3 '-wing consisting of 2 to 5 linked nucleosides; and
a gap between the 5'-wing and the 3'-wing consisting of 6 to 14 linked 2'-deoxynucleosides; and wherein at least one of the 5'-wing and the 3'-wing comprises at least one bicyclic nucleoside; at least one of the 5 '-wing and the 3 '-wing comprises at least one 2 '-substituted nucleoside; and
wherein the nucleobase sequence of the modified oligonucleotide is complementary to the nucleobase
sequence of a target nucleic acid.
2. The compound of claim 1, wherein one of the 5'-wing or the 3'-wing comprises at least one 2'- deoxynucleoside.
3. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-k-(D)8-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
4. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-(D)9-k-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
5. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-k-e-k-(D)9-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
6.
7. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-d-k-d-k-(D)10-e-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
8. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-d-k-d-k-(D)8-e-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
9. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-k-k-(D)7-k-k-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
10. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-d-k-d-k-(D)9-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2 '-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
1 1. The compound of any of claims 1 -2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-e-k-(D)i0-k-e-k, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
12. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-d-k-d-k-(D)8-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
13. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-k-k-(D)8-e-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
14. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: k-e-k-(D)8-e-e-e-e-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
15. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-k-(D)i0-k-e-k-e, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
16. The compound of any of claims 1-2, wherein the compound comprises a modified oligonucleotide having the sugar motif: e-e-e-(D)i0-k-k-k, wherein each k comprises a bicyclic nucleoside, each e comprises a 2'-modified nucleoside, and each D comprises a 2'-deoxynucleoside.
17. A pharmaceutical composition comprising the compound according to any of claims 1-16 and a pharmaceutically acceptable diluent.
18. A method of modulating expression of a target nucleic acid in a cell comprising contacting the cell with a compound according to any of claims 1-16.
19. A method of modulating expression of a target nucleic acid in an animal comprising administering to the animal the pharmaceutical composition according to claim 17.
20. Use of a compound of any of claims 1 to 16 or the pharmaceutical composition of claim 17 for the preparation of a medicament for the treatment of a disease or condition.
PCT/US2013/037638 2012-04-20 2013-04-22 Oligomeric compounds comprising bicyclic nucleotides and uses thereof WO2013159108A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US201261636513P true 2012-04-20 2012-04-20
US61/636,513 2012-04-20

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US14/395,780 US9914922B2 (en) 2012-04-20 2013-04-22 Oligomeric compounds comprising bicyclic nucleotides and uses thereof
EP13777683.7A EP2839006B1 (en) 2012-04-20 2013-04-22 Oligomeric compounds comprising bicyclic nucleotides and uses thereof
US15/880,758 US20180251761A1 (en) 2012-04-20 2018-01-26 Oligomeric compounds comprising bicyclic nucleotides and uses thereof

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US201414395780A A-371-Of-International 2014-10-20 2014-10-20
US15/880,758 Continuation US20180251761A1 (en) 2012-04-20 2018-01-26 Oligomeric compounds comprising bicyclic nucleotides and uses thereof

Publications (2)

Publication Number Publication Date
WO2013159108A2 true WO2013159108A2 (en) 2013-10-24
WO2013159108A3 WO2013159108A3 (en) 2014-01-03

Family

ID=49384239

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2013/037638 WO2013159108A2 (en) 2012-04-20 2013-04-22 Oligomeric compounds comprising bicyclic nucleotides and uses thereof

Country Status (3)

Country Link
US (2) US9914922B2 (en)
EP (2) EP2839006B1 (en)
WO (1) WO2013159108A2 (en)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016040589A1 (en) 2014-09-12 2016-03-17 Alnylam Pharmaceuticals, Inc. Polynucleotide agents targeting complement component c5 and methods of use thereof
WO2016061487A1 (en) 2014-10-17 2016-04-21 Alnylam Pharmaceuticals, Inc. Polynucleotide agents targeting aminolevulinic acid synthase-1 (alas1) and uses thereof
WO2016069694A2 (en) 2014-10-30 2016-05-06 Alnylam Pharmaceuticals, Inc. Polynucleotide agents targeting serpinc1 (at3) and methods of use thereof
WO2016127002A1 (en) * 2015-02-04 2016-08-11 Bristol-Myers Squibb Company Lna oligonucleotides with alternating flanks
WO2016164746A1 (en) 2015-04-08 2016-10-13 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the lect2 gene
WO2016205323A1 (en) 2015-06-18 2016-12-22 Alnylam Pharmaceuticals, Inc. Polynucleotde agents targeting hydroxyacid oxidase (glycolate oxidase, hao1) and methods of use thereof
US9695418B2 (en) 2012-10-11 2017-07-04 Ionis Pharmaceuticals, Inc. Oligomeric compounds comprising bicyclic nucleosides and uses thereof

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2921167A1 (en) 2013-05-01 2014-11-06 Ionis Pharmaceuticals, Inc. Compositions and methods for modulating hbv and ttr expression
US10006027B2 (en) 2014-03-19 2018-06-26 Ionis Pharmaceuticals, Inc. Methods for modulating Ataxin 2 expression
GB201410693D0 (en) 2014-06-16 2014-07-30 Univ Southampton Splicing modulation
CA2963288A1 (en) 2014-10-03 2016-04-07 Cold Spring Harbor Laboratory Targeted augmentation of nuclear gene output
JP2018531600A (en) 2015-10-09 2018-11-01 ユニバーシティ・オブ・サザンプトン Screening of regulation and deregulated protein expression in gene expression

Citations (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3687808A (en) 1969-08-14 1972-08-29 Univ Leland Stanford Junior Synthetic polynucleotides
US4845205A (en) 1985-01-08 1989-07-04 Institut Pasteur 2,N6 -disubstituted and 2,N6 -trisubstituted adenosine-3'-phosphoramidites
US5130302A (en) 1989-12-20 1992-07-14 Boron Bilogicals, Inc. Boronated nucleoside, nucleotide and oligonucleotide compounds, compositions and methods for using same
US5134066A (en) 1989-08-29 1992-07-28 Monsanto Company Improved probes using nucleosides containing 3-dezauracil analogs
US5175273A (en) 1988-07-01 1992-12-29 Genentech, Inc. Nucleic acid intercalating agents
US5367066A (en) 1984-10-16 1994-11-22 Chiron Corporation Oligonucleotides with selectably cleavable and/or abasic sites
US5432272A (en) 1990-10-09 1995-07-11 Benner; Steven A. Method for incorporating into a DNA or RNA oligonucleotide using nucleotides bearing heterocyclic bases
US5457187A (en) 1993-12-08 1995-10-10 Board Of Regents University Of Nebraska Oligonucleotides containing 5-fluorouracil
US5459255A (en) 1990-01-11 1995-10-17 Isis Pharmaceuticals, Inc. N-2 substituted purines
US5484908A (en) 1991-11-26 1996-01-16 Gilead Sciences, Inc. Oligonucleotides containing 5-propynyl pyrimidines
US5502177A (en) 1993-09-17 1996-03-26 Gilead Sciences, Inc. Pyrimidine derivatives for labeled binding partners
US5525711A (en) 1994-05-18 1996-06-11 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Pteridine nucleotide analogs as fluorescent DNA probes
US5552540A (en) 1987-06-24 1996-09-03 Howard Florey Institute Of Experimental Physiology And Medicine Nucleoside derivatives
US5594121A (en) 1991-11-07 1997-01-14 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified purines
US5596091A (en) 1994-03-18 1997-01-21 The Regents Of The University Of California Antisense oligonucleotides comprising 5-aminoalkyl pyrimidine nucleotides
US5614617A (en) 1990-07-27 1997-03-25 Isis Pharmaceuticals, Inc. Nuclease resistant, pyrimidine modified oligonucleotides that detect and modulate gene expression
US5645985A (en) 1991-11-26 1997-07-08 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified pyrimidines
US5681941A (en) 1990-01-11 1997-10-28 Isis Pharmaceuticals, Inc. Substituted purines and oligonucleotide cross-linking
US5750692A (en) 1990-01-11 1998-05-12 Isis Pharmaceuticals, Inc. Synthesis of 3-deazapurines
US5830653A (en) 1991-11-26 1998-11-03 Gilead Sciences, Inc. Methods of using oligomers containing modified pyrimidines
WO2008049085A1 (en) 2006-10-18 2008-04-24 Isis Pharmaceuticals, Inc. Antisense compounds
US7399845B2 (en) 2006-01-27 2008-07-15 Isis Pharmaceuticals, Inc. 6-modified bicyclic nucleic acid analogs
WO2008101157A1 (en) 2007-02-15 2008-08-21 Isis Pharmaceuticals, Inc. 5'-substituted-2'-f modified nucleosides and oligomeric compounds prepared therefrom
WO2011097643A1 (en) 2010-02-08 2011-08-11 Isis Pharmaceuticals, Inc. Selective reduction of allelic variants
WO2012109395A1 (en) 2011-02-08 2012-08-16 Isis Pharmaceuticals, Inc. Oligomeric compounds comprising bicyclic nucleotides and uses thereof

Family Cites Families (85)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2699808A (en) 1944-10-06 1955-01-18 Mark W Lowe Apparatus for peeling tomatoes
US2699508A (en) 1951-12-21 1955-01-11 Selectronics Inc Method of mounting and construction of mounting for low frequency piezoelectric crystals
US5118800A (en) 1983-12-20 1992-06-02 California Institute Of Technology Oligonucleotides possessing a primary amino group in the terminal nucleotide
FR2567892B1 (en) 1984-07-19 1989-02-17 Centre Nat Rech Scient New oligonucleotides, their method of preparing and applications as mediators in the development of the effects of interferons
US6582908B2 (en) 1990-12-06 2003-06-24 Affymetrix, Inc. Oligonucleotides
US5591722A (en) 1989-09-15 1997-01-07 Southern Research Institute 2'-deoxy-4'-thioribonucleosides and their antiviral activity
US5646265A (en) 1990-01-11 1997-07-08 Isis Pharmceuticals, Inc. Process for the preparation of 2'-O-alkyl purine phosphoramidites
US5670633A (en) 1990-01-11 1997-09-23 Isis Pharmaceuticals, Inc. Sugar modified oligonucleotides that detect and modulate gene expression
GB9009980D0 (en) 1990-05-03 1990-06-27 Amersham Int Plc Phosphoramidite derivatives,their preparation and the use thereof in the incorporation of reporter groups on synthetic oligonucleotides
DE69032425D1 (en) 1990-05-11 1998-07-23 Microprobe Corp Test strips for immersion for nucleic acid hybridization assays and methods for the covalent immobilization of oligonucleotides
US20010044145A1 (en) 1991-12-24 2001-11-22 Monia Brett P. Methods of using mammalian RNase H and compositions thereof
ES2103918T3 (en) 1991-10-17 1997-10-01 Ciba Geigy Ag Bicyclic nucleosides, oligonucleotides, process for their preparation and intermediates.
US5359044A (en) 1991-12-13 1994-10-25 Isis Pharmaceuticals Cyclobutyl oligonucleotide surrogates
FR2687679B1 (en) 1992-02-05 1994-10-28 Centre Nat Rech Scient Oligothionucleotides.
FR2692265B1 (en) 1992-05-25 1996-11-08 Centre Nat Rech Scient biologically active compounds of type phosphotriesters.
EP0577558A3 (en) 1992-07-01 1994-04-20 Ciba Geigy Ag
WO1994014226A1 (en) 1992-12-14 1994-06-23 Honeywell Inc. Motor system with individually controlled redundant windings
MX9306994A (en) 1992-12-15 1994-06-30 Ericsson Telefon Ab L M Flow control system for packet switches.
US5446786A (en) 1993-03-03 1995-08-29 Northern Telecom Limited Two-wire telecommunications line detection arrangements
JPH08508492A (en) 1993-03-30 1996-09-10 スターリング ウィンスロップ インコーポレイティド Acyclic nucleoside analogues and oligonucleotide sequences comprising them
DE4311944A1 (en) 1993-04-10 1994-10-13 Degussa Coated sodium percarbonate particles, processes for their preparation and to detergents containing detergent and bleaching compositions
FR2705099B1 (en) 1993-05-12 1995-08-04 Centre Nat Rech Scient Oligonucleotide phosphorothioate triesters and method of preparation.
US5801154A (en) 1993-10-18 1998-09-01 Isis Pharmaceuticals, Inc. Antisense oligonucleotide modulation of multidrug resistance-associated protein
US5446137B1 (en) 1993-12-09 1998-10-06 Behringwerke Ag Oligonucleotides containing 4'-substituted nucleotides
US5519134A (en) 1994-01-11 1996-05-21 Isis Pharmaceuticals, Inc. Pyrrolidine-containing monomers and oligomers
US5627053A (en) 1994-03-29 1997-05-06 Ribozyme Pharmaceuticals, Inc. 2'deoxy-2'-alkylnucleotide containing nucleic acid
US5597909A (en) 1994-08-25 1997-01-28 Chiron Corporation Polynucleotide reagents containing modified deoxyribose moieties, and associated methods of synthesis and use
US5792747A (en) 1995-01-24 1998-08-11 The Administrators Of The Tulane Educational Fund Highly potent agonists of growth hormone releasing hormone
US5656408A (en) 1996-04-29 1997-08-12 Xerox Corporation Coated carrier particles
JP2008501693A (en) 2004-06-03 2008-01-24 アイシス ファーマシューティカルズ、インク. Duplex composition having individually regulated strands for use in gene regulation
US6770748B2 (en) 1997-03-07 2004-08-03 Takeshi Imanishi Bicyclonucleoside and oligonucleotide analogue
JP3756313B2 (en) 1997-03-07 2006-03-15 武 今西 New bicyclo nucleosides and oligonucleotide analogues
WO1999014226A2 (en) 1997-09-12 1999-03-25 Exiqon A/S Bi- and tri-cyclic nucleoside, nucleotide and oligonucleotide analogues
US6794499B2 (en) 1997-09-12 2004-09-21 Exiqon A/S Oligonucleotide analogues
US20030228597A1 (en) 1998-04-13 2003-12-11 Cowsert Lex M. Identification of genetic targets for modulation by oligonucleotides and generation of oligonucleotides for gene modulation
US6043352A (en) 1998-08-07 2000-03-28 Isis Pharmaceuticals, Inc. 2'-O-Dimethylaminoethyloxyethyl-modified oligonucleotides
CN1273478C (en) 1999-02-12 2006-09-06 三共株式会社 Novel nucleosides and oligonucleotide analogues
US7084125B2 (en) 1999-03-18 2006-08-01 Exiqon A/S Xylo-LNA analogues
US7098192B2 (en) 1999-04-08 2006-08-29 Isis Pharmaceuticals, Inc. Antisense oligonucleotide modulation of STAT3 expression
ES2283298T3 (en) 1999-05-04 2007-11-01 Santaris Pharma A/S Analogs l-ribo-LNA.
US6525191B1 (en) 1999-05-11 2003-02-25 Kanda S. Ramasamy Conformationally constrained L-nucleosides
US20040002153A1 (en) 1999-07-21 2004-01-01 Monia Brett P. Modulation of PTEN expression via oligomeric compounds
JP4151751B2 (en) 1999-07-22 2008-09-17 第一三共株式会社 New bicycloalkyl nucleoside analogues
AU2002317437A1 (en) 2001-05-18 2002-12-03 Cureon A/S Therapeutic uses of lna-modified oligonucleotides in infectious diseases
US7888324B2 (en) 2001-08-01 2011-02-15 Genzyme Corporation Antisense modulation of apolipoprotein B expression
US7569575B2 (en) 2002-05-08 2009-08-04 Santaris Pharma A/S Synthesis of locked nucleic acid derivatives
JP4986109B2 (en) 2002-11-13 2012-07-25 ジェンザイム・コーポレーション Antisense modulation of apolipoprotein b expression
US20040219565A1 (en) 2002-10-21 2004-11-04 Sakari Kauppinen Oligonucleotides useful for detecting and analyzing nucleic acids of interest
WO2004043979A2 (en) 2002-11-05 2004-05-27 Isis Pharmaceuticals, Inc. Sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
EP1562971B1 (en) 2002-11-05 2014-02-12 Isis Pharmaceuticals, Inc. Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
DE60329220D1 (en) 2002-11-18 2009-10-22 Santaris Pharma As Antisense design
EP1592793B2 (en) 2003-02-10 2014-05-07 Santaris Pharma A/S Oligomeric compounds for the modulation of survivin expression
WO2004106356A1 (en) 2003-05-27 2004-12-09 Syddansk Universitet Functionalized nucleotide derivatives
US7427672B2 (en) 2003-08-28 2008-09-23 Takeshi Imanishi Artificial nucleic acids of n-o bond crosslinkage type
US20050074801A1 (en) 2003-09-09 2005-04-07 Monia Brett P. Chimeric oligomeric compounds comprising alternating regions of northern and southern conformational geometry
US20050053981A1 (en) 2003-09-09 2005-03-10 Swayze Eric E. Gapped oligomeric compounds having linked bicyclic sugar moieties at the termini
EP1677822B1 (en) 2003-09-18 2014-04-23 Isis Pharmaceuticals, Inc. 4'-thionucleosides and oligomeric compounds
CN100558893C (en) 2003-09-18 2009-11-11 Isis 药物公司;伊莱利利公司 Modulation of eIF4E expression
AT467679T (en) 2003-12-23 2010-05-15 Santaris Pharma As Oligomeric compounds for the modulation of bcl-2
GB0407382D0 (en) 2004-03-31 2004-05-05 Univ Cambridge Tech Therapeutic methods and means
JP5523705B2 (en) 2005-08-29 2014-06-18 レグルス・セラピューティクス・インコーポレイテッドRegulus Therapeutics Inc. Mir-122a how to use to modulate
WO2007027894A2 (en) 2005-08-29 2007-03-08 Isis Pharmaceuticals, Inc. Antisense compounds having enhanced anti-microrna activity
PT2161038E (en) 2006-01-26 2014-03-10 Isis Pharmaceuticals Inc Compositions and their uses directed to huntingtin
WO2007134014A2 (en) 2006-05-05 2007-11-22 Isis Pharmaceuticals, Inc. Compounds and methods for modulating expression of gcgr
ES2389737T3 (en) 2006-05-11 2012-10-31 Isis Pharmaceuticals, Inc. Bicyclic analogues modified nucleic acids 5 '
US8461124B2 (en) 2007-03-15 2013-06-11 Jyoti Chattopadhyaya Five- and six-membered conformationally locked 2′,4′-carbocyclic ribo-thymidines for the treatment of infections and cancer
EP2170917B1 (en) 2007-05-30 2012-06-27 Isis Pharmaceuticals, Inc. N-substituted-aminomethylene bridged bicyclic nucleic acid analogs
EP2173760B2 (en) 2007-06-08 2015-11-04 Isis Pharmaceuticals, Inc. Carbocyclic bicyclic nucleic acid analogs
CA2692579C (en) 2007-07-05 2016-05-03 Isis Pharmaceuticals, Inc. 6-disubstituted bicyclic nucleic acid analogs
CN101821277B (en) 2007-08-15 2014-05-07 Isis制药公司 Tetrahydropyran nucleic acid analogs
WO2009061841A2 (en) 2007-11-05 2009-05-14 Isis Pharmaceuticals, Inc. Modified polynucleotides as antidotes to antisense compounds
US9029337B2 (en) 2007-11-09 2015-05-12 Isis Pharmaceuticals, Inc. Modulation of factor 7 expression
WO2009067647A1 (en) 2007-11-21 2009-05-28 Isis Pharmaceuticals, Inc. Carbocyclic alpha-l-bicyclic nucleic acid analogs
US8530640B2 (en) 2008-02-07 2013-09-10 Isis Pharmaceuticals, Inc. Bicyclic cyclohexitol nucleic acid analogs
EP2274423A2 (en) * 2008-04-04 2011-01-19 Isis Pharmaceuticals, Inc. Oligomeric compounds comprising bicyclic nucleosides and having reduced toxicity
EP2356129B1 (en) 2008-09-24 2013-04-03 Isis Pharmaceuticals, Inc. Substituted alpha-l-bicyclic nucleosides
AT507215B1 (en) 2009-01-14 2010-03-15 Boehler Edelstahl Gmbh & Co Kg Verschleissbeständiger material
WO2010108035A1 (en) 2009-03-18 2010-09-23 Isis Pharmaceuticals, Inc. Compounds and methods for modulating toxic and proinflammatory effects
WO2011017521A2 (en) 2009-08-06 2011-02-10 Isis Pharmaceuticals, Inc. Bicyclic cyclohexose nucleic acid analogs
AU2011213563B2 (en) 2010-02-08 2015-12-24 Ionis Pharmaceuticals, Inc. Selective reduction of allelic variants
WO2012027033A1 (en) 2010-07-19 2012-03-01 Isis Pharmaceuticals, Inc. Compounds and methods for modulating target nuclear and sub-nuclear nucleic acid molecules in cells and animals
EP2699583A4 (en) 2011-04-21 2015-04-15 Isis Pharmaceuticals Inc Modulation of hepatitis b virus (hbv) expression
EP2742136B1 (en) 2011-08-11 2017-09-27 Ionis Pharmaceuticals, Inc. Gapped oligomeric compounds comprising 5'-modified deoxyribonucleosides in the gap and uses thereof
US9984408B1 (en) 2012-05-30 2018-05-29 Amazon Technologies, Inc. Method, medium, and system for live video cooperative shopping
US9778708B1 (en) 2016-07-18 2017-10-03 Lenovo Enterprise Solutions (Singapore) Pte. Ltd. Dual sided latching retainer for computer modules

Patent Citations (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3687808A (en) 1969-08-14 1972-08-29 Univ Leland Stanford Junior Synthetic polynucleotides
US5367066A (en) 1984-10-16 1994-11-22 Chiron Corporation Oligonucleotides with selectably cleavable and/or abasic sites
US4845205A (en) 1985-01-08 1989-07-04 Institut Pasteur 2,N6 -disubstituted and 2,N6 -trisubstituted adenosine-3'-phosphoramidites
US5552540A (en) 1987-06-24 1996-09-03 Howard Florey Institute Of Experimental Physiology And Medicine Nucleoside derivatives
US5175273A (en) 1988-07-01 1992-12-29 Genentech, Inc. Nucleic acid intercalating agents
US5134066A (en) 1989-08-29 1992-07-28 Monsanto Company Improved probes using nucleosides containing 3-dezauracil analogs
US5130302A (en) 1989-12-20 1992-07-14 Boron Bilogicals, Inc. Boronated nucleoside, nucleotide and oligonucleotide compounds, compositions and methods for using same
US5587469A (en) 1990-01-11 1996-12-24 Isis Pharmaceuticals, Inc. Oligonucleotides containing N-2 substituted purines
US5459255A (en) 1990-01-11 1995-10-17 Isis Pharmaceuticals, Inc. N-2 substituted purines
US5681941A (en) 1990-01-11 1997-10-28 Isis Pharmaceuticals, Inc. Substituted purines and oligonucleotide cross-linking
US5750692A (en) 1990-01-11 1998-05-12 Isis Pharmaceuticals, Inc. Synthesis of 3-deazapurines
US5614617A (en) 1990-07-27 1997-03-25 Isis Pharmaceuticals, Inc. Nuclease resistant, pyrimidine modified oligonucleotides that detect and modulate gene expression
US5432272A (en) 1990-10-09 1995-07-11 Benner; Steven A. Method for incorporating into a DNA or RNA oligonucleotide using nucleotides bearing heterocyclic bases
US5594121A (en) 1991-11-07 1997-01-14 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified purines
US5830653A (en) 1991-11-26 1998-11-03 Gilead Sciences, Inc. Methods of using oligomers containing modified pyrimidines
US5645985A (en) 1991-11-26 1997-07-08 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified pyrimidines
US5484908A (en) 1991-11-26 1996-01-16 Gilead Sciences, Inc. Oligonucleotides containing 5-propynyl pyrimidines
US6005096A (en) 1993-09-17 1999-12-21 Gilead Sciences, Inc. Pyrimidine derivatives
US5502177A (en) 1993-09-17 1996-03-26 Gilead Sciences, Inc. Pyrimidine derivatives for labeled binding partners
US5763588A (en) 1993-09-17 1998-06-09 Gilead Sciences, Inc. Pyrimidine derivatives for labeled binding partners
US5457187A (en) 1993-12-08 1995-10-10 Board Of Regents University Of Nebraska Oligonucleotides containing 5-fluorouracil
US5596091A (en) 1994-03-18 1997-01-21 The Regents Of The University Of California Antisense oligonucleotides comprising 5-aminoalkyl pyrimidine nucleotides
US5525711A (en) 1994-05-18 1996-06-11 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Pteridine nucleotide analogs as fluorescent DNA probes
US7399845B2 (en) 2006-01-27 2008-07-15 Isis Pharmaceuticals, Inc. 6-modified bicyclic nucleic acid analogs
WO2008049085A1 (en) 2006-10-18 2008-04-24 Isis Pharmaceuticals, Inc. Antisense compounds
WO2008101157A1 (en) 2007-02-15 2008-08-21 Isis Pharmaceuticals, Inc. 5'-substituted-2'-f modified nucleosides and oligomeric compounds prepared therefrom
WO2011097643A1 (en) 2010-02-08 2011-08-11 Isis Pharmaceuticals, Inc. Selective reduction of allelic variants
WO2012109395A1 (en) 2011-02-08 2012-08-16 Isis Pharmaceuticals, Inc. Oligomeric compounds comprising bicyclic nucleotides and uses thereof

Non-Patent Citations (21)

* Cited by examiner, † Cited by third party
Title
"Carbohydrate Modifications in Antisense Research", ACS SYMPOSIUM SERIES 580, pages: 40 - 65
"Strategies, and Applications", CRC PRESS
"The Concise Encyclopedia Of Polymer Science And Engineering", 1990, JOHN WILEY & SONS, pages: 858 - 859
CROOKE ET AL., J. PHARMACOL. EXP. THER., vol. 277, 1996, pages 923 - 937
ENGLISCH ET AL.: "Angewandte Chemie", vol. 30, 1991, pages: 613
KABANOV ET AL., FEBS LETT., vol. 259, 1990, pages 327 - 330
LETSINGER ET AL., PROC. NATL. ACAD. SCI. USA, vol. 86, 1989, pages 6553 - 6556
MANOHARAN ET AL., ANN. N.Y. ACAD. SCI., vol. 660, 1992, pages 306 - 309
MANOHARAN ET AL., BIOORG. MED. CHEM. LET., vol. 3, 1993, pages 2765 - 2770
MANOHARAN ET AL., BIOORG. MED. CHEM. LET., vol. 4, 1994, pages 1053 - 1060
MANOHARAN ET AL., NUCLEOSIDES & NUCLEOTIDES, vol. 14, 1995, pages 969 - 973
MANOHARAN ET AL., TETRAHEDRON LETT., vol. 36, 1995, pages 3651 - 3654
MISHRA ET AL., BIOCHIM. BIOPHYS. ACTA, vol. 1264, 1995, pages 229 - 237
OBERHAUSER ET AL., NUCL. ACIDS RES., vol. 20, 1992, pages 533 - 538
SAISON-BEHMOARAS ET AL., EMBO J., vol. 10, 1991, pages 1111 - 1118
SAMBROOK ET AL.: "Molecular Cloning, A laboratory Manual", 1989, COLD SPRING HARBOR LABORATORY PRESS
SANGHVI, Y.S.: "Antisense Research and Applications", 1993, CRC PRESS, pages: 273 - 288
See also references of EP2839006A4
SHEA ET AL., NUCL. ACIDS RES., vol. 18, 1990, pages 3777 - 3783
SVINARCHUK ET AL., BIOCHIMIE, vol. 75, 1993, pages 49 - 54
W. F. LIMA: "The Positional Influence of the Helical Geometry of the Heteroduplex Substrate on Human RNase HI Catalysis", MOLECULAR PHARMACOLOGY, vol. 71, no. 1, 2007, pages 73 - 82, XP055225537, DOI: doi:10.1124/mol.106.025429

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9695418B2 (en) 2012-10-11 2017-07-04 Ionis Pharmaceuticals, Inc. Oligomeric compounds comprising bicyclic nucleosides and uses thereof
WO2016040589A1 (en) 2014-09-12 2016-03-17 Alnylam Pharmaceuticals, Inc. Polynucleotide agents targeting complement component c5 and methods of use thereof
WO2016061487A1 (en) 2014-10-17 2016-04-21 Alnylam Pharmaceuticals, Inc. Polynucleotide agents targeting aminolevulinic acid synthase-1 (alas1) and uses thereof
WO2016069694A2 (en) 2014-10-30 2016-05-06 Alnylam Pharmaceuticals, Inc. Polynucleotide agents targeting serpinc1 (at3) and methods of use thereof
WO2016069694A3 (en) * 2014-10-30 2016-06-23 Alnylam Pharmaceuticals, Inc. Polynucleotide agents targeting serpinc1 (at3) and methods of use thereof
WO2016127002A1 (en) * 2015-02-04 2016-08-11 Bristol-Myers Squibb Company Lna oligonucleotides with alternating flanks
WO2016164746A1 (en) 2015-04-08 2016-10-13 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the lect2 gene
WO2016205323A1 (en) 2015-06-18 2016-12-22 Alnylam Pharmaceuticals, Inc. Polynucleotde agents targeting hydroxyacid oxidase (glycolate oxidase, hao1) and methods of use thereof

Also Published As

Publication number Publication date
US20150184153A1 (en) 2015-07-02
US20180251761A1 (en) 2018-09-06
US9914922B2 (en) 2018-03-13
EP2839006B1 (en) 2018-01-03
EP3336189A1 (en) 2018-06-20
EP2839006A4 (en) 2015-12-16
EP2839006A2 (en) 2015-02-25
WO2013159108A3 (en) 2014-01-03

Similar Documents

Publication Publication Date Title
AU2017203436B2 (en) Compositions and methods for modulating apolipoprotein c-iii expression
EP2458006B1 (en) Compounds and methods for modulating expression APOB
US9765338B2 (en) Modulation of dystrophia myotonica-protein kinase (DMPK) expression
EP2920307B1 (en) Anti apob antisense conjugate compounds
EP2444494B1 (en) Rna antagonist compounds for the modulation of pcsk9
US8440637B2 (en) Combination treatment for the treatment of hepatitis C virus infection
RU2649367C2 (en) Lna oligonucleotide carbohydrate conjugates
JP5665317B2 (en) Antisense compounds
EP2601204A2 (en) Modified nucleosides and oligomeric compounds prepared therefrom
AU2006291836A1 (en) RNA antagonist compounds for the inhibition of Apo-Bl00 expression
US20150018540A1 (en) Oligomer-conjugate complexes and their use
US20130059902A1 (en) Methods and compositions useful in treatment of diseases or conditions related to repeat expansion
KR101931089B1 (en) Antisense oligomers and conjugates targeting pcsk9
ES2651514T3 (en) oligomeric compounds comprising deoxirribonucleósidos gapados modified 5&#39;-in the gap and uses thereof
US8846639B2 (en) Oligomeric compounds comprising bicyclic nucleosides and having reduced toxicity
WO2010048585A2 (en) Oligomeric compounds and methods
US10179912B2 (en) RNA modulating oligonucleotides with improved characteristics for the treatment of duchenne and becker muscular dystrophy
WO2014118272A1 (en) Antimir-122 oligonucleotide carbohydrate conjugates
US9399774B2 (en) Modulation of transthyretin expression
EP2310505A1 (en) Antidote oligomers
US8563528B2 (en) Antisense oligomers targeting PCSK9
US20180179521A1 (en) Modified crispr rna and modified single crispr rna and uses thereof
AU2013253074B2 (en) RNA modulating oligonucleotides with improved characteristics for the treatment of neuromuscular disorders
EP2812342B1 (en) Modulation of rna by repeat targeting
JP2017501684A (en) Apob antisense conjugate compound

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13777683

Country of ref document: EP

Kind code of ref document: A2

WWE Wipo information: entry into national phase

Ref document number: 14395780

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 2013777683

Country of ref document: EP

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13777683

Country of ref document: EP

Kind code of ref document: A2